human colorectal cancer cell lines  (ATCC)


Bioz Verified Symbol ATCC is a verified supplier
Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    ATCC human colorectal cancer cell lines
    Endogenous expression of Pdcd4, CD24, Src, miR-21 and miR-34a in resected <t>colorectal</t> tissues. ( a ) Western blot analysis was performed for Pdcd4, CD24 and Src in colorectal tumors (Tumor) and corresponding normal tissues (Normal) taken from a series of 26 patients. β-Actin served as internal control. Relative mean protein amounts (Fold change comparative to normal tissue expression) of Pdcd4, CD24 and Src obtained by densitometry analysis are represented as bar graphs. Specific Pdcd4, CD24 or Src band intersities were normalized with β-actin. Pdcd4 was downregulated, CD24 and Src were upregulated significantly in the tumor tissues (p = 0.003, p = 0.05 and p = 0.001, respectively) ( b ) Real-time PCR results of miR-21 and miR-34a in the same colorectal tumor (Tumor) and normal tissue (Normal) samples. Mean relative expression (fold change compared to expression in normal tissue) of miR-21 and miR-34a is represented as bar graphs. miR-21 was upregulated and miR-34a was downregulated significantly in the tumor tissues. (p = 0.002, p = 0.05, respectively) ( c ) Lysates from 7 representative normal tissue (N) and colorectal tumor (T) samples were subjected to Western blotting and probed for the expression of Pdcd4, CD24 and Src and represented. β-Actin served as a loading control ( d ) Schematic representation of the functional network between CD24, Src, AP-1, miR-21, Pdcd4 and miR-34a.
    Human Colorectal Cancer Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human colorectal cancer cell lines/product/ATCC
    Average 86 stars, based on 40 article reviews
    Price from $9.99 to $1999.99
    human colorectal cancer cell lines - by Bioz Stars, 2022-09
    86/100 stars

    Images

    1) Product Images from "CD24 Induces Expression of the Oncomir miR-21 via Src, and CD24 and Src Are Both Post-Transcriptionally Downregulated by the Tumor Suppressor miR-34a"

    Article Title: CD24 Induces Expression of the Oncomir miR-21 via Src, and CD24 and Src Are Both Post-Transcriptionally Downregulated by the Tumor Suppressor miR-34a

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0059563

    Endogenous expression of Pdcd4, CD24, Src, miR-21 and miR-34a in resected colorectal tissues. ( a ) Western blot analysis was performed for Pdcd4, CD24 and Src in colorectal tumors (Tumor) and corresponding normal tissues (Normal) taken from a series of 26 patients. β-Actin served as internal control. Relative mean protein amounts (Fold change comparative to normal tissue expression) of Pdcd4, CD24 and Src obtained by densitometry analysis are represented as bar graphs. Specific Pdcd4, CD24 or Src band intersities were normalized with β-actin. Pdcd4 was downregulated, CD24 and Src were upregulated significantly in the tumor tissues (p = 0.003, p = 0.05 and p = 0.001, respectively) ( b ) Real-time PCR results of miR-21 and miR-34a in the same colorectal tumor (Tumor) and normal tissue (Normal) samples. Mean relative expression (fold change compared to expression in normal tissue) of miR-21 and miR-34a is represented as bar graphs. miR-21 was upregulated and miR-34a was downregulated significantly in the tumor tissues. (p = 0.002, p = 0.05, respectively) ( c ) Lysates from 7 representative normal tissue (N) and colorectal tumor (T) samples were subjected to Western blotting and probed for the expression of Pdcd4, CD24 and Src and represented. β-Actin served as a loading control ( d ) Schematic representation of the functional network between CD24, Src, AP-1, miR-21, Pdcd4 and miR-34a.
    Figure Legend Snippet: Endogenous expression of Pdcd4, CD24, Src, miR-21 and miR-34a in resected colorectal tissues. ( a ) Western blot analysis was performed for Pdcd4, CD24 and Src in colorectal tumors (Tumor) and corresponding normal tissues (Normal) taken from a series of 26 patients. β-Actin served as internal control. Relative mean protein amounts (Fold change comparative to normal tissue expression) of Pdcd4, CD24 and Src obtained by densitometry analysis are represented as bar graphs. Specific Pdcd4, CD24 or Src band intersities were normalized with β-actin. Pdcd4 was downregulated, CD24 and Src were upregulated significantly in the tumor tissues (p = 0.003, p = 0.05 and p = 0.001, respectively) ( b ) Real-time PCR results of miR-21 and miR-34a in the same colorectal tumor (Tumor) and normal tissue (Normal) samples. Mean relative expression (fold change compared to expression in normal tissue) of miR-21 and miR-34a is represented as bar graphs. miR-21 was upregulated and miR-34a was downregulated significantly in the tumor tissues. (p = 0.002, p = 0.05, respectively) ( c ) Lysates from 7 representative normal tissue (N) and colorectal tumor (T) samples were subjected to Western blotting and probed for the expression of Pdcd4, CD24 and Src and represented. β-Actin served as a loading control ( d ) Schematic representation of the functional network between CD24, Src, AP-1, miR-21, Pdcd4 and miR-34a.

    Techniques Used: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Functional Assay

    2) Product Images from "MicroRNA-628-5p inhibits cell proliferation and induces apoptosis in colorectal cancer through downregulating CCND1 expression levels"

    Article Title: MicroRNA-628-5p inhibits cell proliferation and induces apoptosis in colorectal cancer through downregulating CCND1 expression levels

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2020.10945

    miR-6285-p expression levels are decreased in colorectal cancer in vivo and in vitro . (A and B) The expression levels of miR-628-5p were detected using reverse transcription-quantitative PCR in (A) colorectal cancer tissues and (B) colorectal cancer cell lines. **P
    Figure Legend Snippet: miR-6285-p expression levels are decreased in colorectal cancer in vivo and in vitro . (A and B) The expression levels of miR-628-5p were detected using reverse transcription-quantitative PCR in (A) colorectal cancer tissues and (B) colorectal cancer cell lines. **P

    Techniques Used: Expressing, In Vivo, In Vitro, Real-time Polymerase Chain Reaction

    3) Product Images from "Hormetic dose response to L-ascorbic acid as an anti-cancer drug in colorectal cancer cell lines according to SVCT-2 expression"

    Article Title: Hormetic dose response to L-ascorbic acid as an anti-cancer drug in colorectal cancer cell lines according to SVCT-2 expression

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-29386-7

    Hormetic response with gradient L -ascorbic acid treatment in low SVCT-2 expressing cell lines but not in high SVCT-2 expressing cell lines. ( A ) Cell viability of high SVCT-2 expressing cell lines with gradient L -ascorbic acid treatment. ( B ) Cell viability of low SVCT-2 expressing cell lines with gradient L -ascorbic acid treatment. ( C ) Clustering and heat-map visualization of the response of eight colorectal cancer cell lines to L -ascorbic acid. Drug sensitivity is presented by the area under curve (AUC), and the hormetic responsiveness index, a statistical estimate of the response pattern.
    Figure Legend Snippet: Hormetic response with gradient L -ascorbic acid treatment in low SVCT-2 expressing cell lines but not in high SVCT-2 expressing cell lines. ( A ) Cell viability of high SVCT-2 expressing cell lines with gradient L -ascorbic acid treatment. ( B ) Cell viability of low SVCT-2 expressing cell lines with gradient L -ascorbic acid treatment. ( C ) Clustering and heat-map visualization of the response of eight colorectal cancer cell lines to L -ascorbic acid. Drug sensitivity is presented by the area under curve (AUC), and the hormetic responsiveness index, a statistical estimate of the response pattern.

    Techniques Used: Expressing

    SVCT-2 expression, cytotoxicity, and uptake of L -ascorbic acid in colorectal cancer cell lines. ( A ) SVCT-2 expression was analyzed by western blotting. GAPDH was used as a loading control. ( B ) HPLC analysis of the uptake of L -ascorbic acid. ( C ) Relative SVCT-2 expression determined with Image J program analysis (black bars) and cell viability with 1 mM L -ascorbic acid treatment (white bars).
    Figure Legend Snippet: SVCT-2 expression, cytotoxicity, and uptake of L -ascorbic acid in colorectal cancer cell lines. ( A ) SVCT-2 expression was analyzed by western blotting. GAPDH was used as a loading control. ( B ) HPLC analysis of the uptake of L -ascorbic acid. ( C ) Relative SVCT-2 expression determined with Image J program analysis (black bars) and cell viability with 1 mM L -ascorbic acid treatment (white bars).

    Techniques Used: Expressing, Western Blot, High Performance Liquid Chromatography

    4) Product Images from "Primary tumor- and metastasis-derived colon cancer cells differently modulate connexin expression and function in human capillary endothelial cells"

    Article Title: Primary tumor- and metastasis-derived colon cancer cells differently modulate connexin expression and function in human capillary endothelial cells

    Journal: Oncotarget

    doi:

    Hypothetical model of the endothelial connexin contribution to the colorectal cancer (CRC) pathogenesis The diagram shows the endothelial cell (EC) expression of both Cx32 and Cx43 as well as their ability to form hemi-channels or gap junction channels with CRC cells at the microvascular level. Cancer cells from a primary tumor (here, SW480 cells) locally invade the surrounding tissue, enter the microvasculature of the blood system (passive intravasation), survive and translocate through the bloodstream to micro-vessels of distant tissues. SW480 cells release HSP27 that favors the establishment of GJIC, via Cx43-channels, with the underlying endothelium. This direct cell-to-cell communication contributes to their trans-endothelial migration TEM (active extravasation). In contrast, cancer cells from a metastatic site (here, SW620 cells) release larger amount of chemokines, increasing the endothelial expression of the receptor CXCR2. In turn, CXCR2 promotes both endothelial Cx32 expression and tubulogenesis. The release of ATP through Cx32 hemi-channels from ECs and the subsequent ATP-mediated activation of purinergic P2Y2 receptors could modulate crosstalk between ECs and metastatic cancer cells, favoring neo-angiogenesis in metastatic foci.
    Figure Legend Snippet: Hypothetical model of the endothelial connexin contribution to the colorectal cancer (CRC) pathogenesis The diagram shows the endothelial cell (EC) expression of both Cx32 and Cx43 as well as their ability to form hemi-channels or gap junction channels with CRC cells at the microvascular level. Cancer cells from a primary tumor (here, SW480 cells) locally invade the surrounding tissue, enter the microvasculature of the blood system (passive intravasation), survive and translocate through the bloodstream to micro-vessels of distant tissues. SW480 cells release HSP27 that favors the establishment of GJIC, via Cx43-channels, with the underlying endothelium. This direct cell-to-cell communication contributes to their trans-endothelial migration TEM (active extravasation). In contrast, cancer cells from a metastatic site (here, SW620 cells) release larger amount of chemokines, increasing the endothelial expression of the receptor CXCR2. In turn, CXCR2 promotes both endothelial Cx32 expression and tubulogenesis. The release of ATP through Cx32 hemi-channels from ECs and the subsequent ATP-mediated activation of purinergic P2Y2 receptors could modulate crosstalk between ECs and metastatic cancer cells, favoring neo-angiogenesis in metastatic foci.

    Techniques Used: Expressing, Migration, Transmission Electron Microscopy, Activation Assay

    SW620 cell-secreted factors overexpress the endothelial Cx32 favoring tubulogenesis A. Endothelial cell localization of Cx32 in CRC cell-conditioned media. HMEC were stimulated with SW480-CM or SW620-CM for 6 h and double-stained for ZO-1 and Cx32. Representative micrographs showing the strong labelling of Cx32 induced by SW620-CM and the combined image of co-localization with ZO-1 (yellow); DAPI staining of nuclei ( n = 3, bar 20 μm). B. SW620-CM increase the Cx32 expression in HMEC. A higher Cx32 protein level was detected in response to SW620-CM compared with SW480-CM by immune-blot analysis (no cell expression in unstimulated HMEC). Representative of 5 experiments (Hsc70 as loading control; 150 μg/lane). C-D. In vitro tubulogenesis assay of HMEC pretreated or not (control IgG) with inhibitory monoclonal antibody against Cx32 (anti-Cx32 mAb). HMEC were plated on Matrigel-coated 24-well plates, incubated with SW620-CM for 6 h, and photographed. C. Representative photos of tube formation in HMEC intracellularly delivered with 0.2 μg anti-Cx32 mAb or control IgG (Bar 80 μm). The dotted areas are enlarged in the inserts on the right. Arrows indicated branch points. D. Number of branch points per field of view was quantified (at least 80 single cells were scored; mean ± SD, n = 4; * P
    Figure Legend Snippet: SW620 cell-secreted factors overexpress the endothelial Cx32 favoring tubulogenesis A. Endothelial cell localization of Cx32 in CRC cell-conditioned media. HMEC were stimulated with SW480-CM or SW620-CM for 6 h and double-stained for ZO-1 and Cx32. Representative micrographs showing the strong labelling of Cx32 induced by SW620-CM and the combined image of co-localization with ZO-1 (yellow); DAPI staining of nuclei ( n = 3, bar 20 μm). B. SW620-CM increase the Cx32 expression in HMEC. A higher Cx32 protein level was detected in response to SW620-CM compared with SW480-CM by immune-blot analysis (no cell expression in unstimulated HMEC). Representative of 5 experiments (Hsc70 as loading control; 150 μg/lane). C-D. In vitro tubulogenesis assay of HMEC pretreated or not (control IgG) with inhibitory monoclonal antibody against Cx32 (anti-Cx32 mAb). HMEC were plated on Matrigel-coated 24-well plates, incubated with SW620-CM for 6 h, and photographed. C. Representative photos of tube formation in HMEC intracellularly delivered with 0.2 μg anti-Cx32 mAb or control IgG (Bar 80 μm). The dotted areas are enlarged in the inserts on the right. Arrows indicated branch points. D. Number of branch points per field of view was quantified (at least 80 single cells were scored; mean ± SD, n = 4; * P

    Techniques Used: Staining, Expressing, In Vitro, Incubation

    SW620 cell-secreted factors require CXCR2 signaling pathway to induce the endothelial Cx32 expression and tube formation A. IL-8 secretion in conditioned media from SW480 and SW620 cells was examined through ELISA. CRC cells were exposed or not to the HMEC-CM. All cell media were collected after 6 h (mean ± SD, * P -values
    Figure Legend Snippet: SW620 cell-secreted factors require CXCR2 signaling pathway to induce the endothelial Cx32 expression and tube formation A. IL-8 secretion in conditioned media from SW480 and SW620 cells was examined through ELISA. CRC cells were exposed or not to the HMEC-CM. All cell media were collected after 6 h (mean ± SD, * P -values

    Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay

    HSP27 knockdown inhibits the gap junctional coupling between SW480 cells and HMEC A. Expression of HSP27 in the two CRC cell lines, SW480 and SW620 cells. Detectable amounts of HSP27 in supernatants of SW480 cells but not of SW620 cells (media collected after 12 h). Immunoblots representative of 5 experiments (Hsc70 as loading control). Values indicate amounts of HSP27 measured by ELISA in supernatant of SW480 and SW620 cells for 12 h (mean ± SD; n = 4; P -values
    Figure Legend Snippet: HSP27 knockdown inhibits the gap junctional coupling between SW480 cells and HMEC A. Expression of HSP27 in the two CRC cell lines, SW480 and SW620 cells. Detectable amounts of HSP27 in supernatants of SW480 cells but not of SW620 cells (media collected after 12 h). Immunoblots representative of 5 experiments (Hsc70 as loading control). Values indicate amounts of HSP27 measured by ELISA in supernatant of SW480 and SW620 cells for 12 h (mean ± SD; n = 4; P -values

    Techniques Used: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

    5) Product Images from "Nodal Promotes the Self-Renewal of Human Colon Cancer Stem Cells via an Autocrine Manner through Smad2/3 Signaling Pathway"

    Article Title: Nodal Promotes the Self-Renewal of Human Colon Cancer Stem Cells via an Autocrine Manner through Smad2/3 Signaling Pathway

    Journal: BioMed Research International

    doi: 10.1155/2014/364134

    Protein expression of Nodal ligand and its receptors in various human colorectal cancer cell lines. (a–c) Immunocytochemistry showed translation of NODAL (a), ALK-4 (b), and ACTR-IIB (c) in human colon cancer cell lines, including SW480 cells, LOVO cells, and HCT116 cells. Scale bars in (a)–(c) = 10 μ m.
    Figure Legend Snippet: Protein expression of Nodal ligand and its receptors in various human colorectal cancer cell lines. (a–c) Immunocytochemistry showed translation of NODAL (a), ALK-4 (b), and ACTR-IIB (c) in human colon cancer cell lines, including SW480 cells, LOVO cells, and HCT116 cells. Scale bars in (a)–(c) = 10 μ m.

    Techniques Used: Expressing, Immunocytochemistry

    mRNA expression of Nodal ligand and its three receptors in various human colorectal cancer cell lines. (a–c) RT-PCR revealed the transcripts of Nodal, ALK-4, ALK-7, and Actr-IIb in human colon cancer cell lines, including SW480 cells (a), LOVO cells (b), and HCT116 cells (c). Gapdh served as a loading control of total RNA.
    Figure Legend Snippet: mRNA expression of Nodal ligand and its three receptors in various human colorectal cancer cell lines. (a–c) RT-PCR revealed the transcripts of Nodal, ALK-4, ALK-7, and Actr-IIb in human colon cancer cell lines, including SW480 cells (a), LOVO cells (b), and HCT116 cells (c). Gapdh served as a loading control of total RNA.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    6) Product Images from "GLUT3 Promotes Epithelial–Mesenchymal Transition via TGF-β/JNK/ATF2 Signaling Pathway in Colorectal Cancer Cells"

    Article Title: GLUT3 Promotes Epithelial–Mesenchymal Transition via TGF-β/JNK/ATF2 Signaling Pathway in Colorectal Cancer Cells

    Journal: Biomedicines

    doi: 10.3390/biomedicines10081837

    Schematic representation of the regulation of GLUT3 on TGF-β-mediated EMT in CRC cells. Expression of GLUT3 is regulated by the TGF-β/JNK/ATF2 signaling pathways, and overexpression of GLUT3, thereby, exacerbates CRC cells’ invasiveness. “↓” indicates induction; “├” indicates inhibition. GLUT3, glucose transporter 3; TGF-β, transforming growth factor-β; EMT, epithelial–mesenchymal transition; CRC, colorectal cancer; JNK, c-Jun N-terminal kinase; ATF2, activating transcription factor-2.
    Figure Legend Snippet: Schematic representation of the regulation of GLUT3 on TGF-β-mediated EMT in CRC cells. Expression of GLUT3 is regulated by the TGF-β/JNK/ATF2 signaling pathways, and overexpression of GLUT3, thereby, exacerbates CRC cells’ invasiveness. “↓” indicates induction; “├” indicates inhibition. GLUT3, glucose transporter 3; TGF-β, transforming growth factor-β; EMT, epithelial–mesenchymal transition; CRC, colorectal cancer; JNK, c-Jun N-terminal kinase; ATF2, activating transcription factor-2.

    Techniques Used: Expressing, Over Expression, Inhibition

    Overexpression and knockdown of GLUT3 regulates stemness markers in CRC cells. ( A ) HCT116 cells were transfected with GLUT3/pEJ-3HA (2 µg) for 24 h. Western blot analysis was conducted for measuring the protein levels of GLUT3, Nanog and OCT3/4. ( B ) qRT-PCR was conducted to measure the mRNA levels of GLUT3, Bmi1, Nanog and OCT3/4. ( C ) Western blot analysis was conducted for measuring the protein levels of GLUT3, Nanog and OCT3/4 in shGLUT3-transfected SW620 cells. ( D ) qRT-PCR was conducted to measure the mRNA levels of GLUT3, Bmi1, Nanog and OCT3/4 in shGLUT3-transfected SW620 cells. GLUT3, glucose transporter 3; CRC, colorectal cancer; qRT-PCR, quantitative reverse transcription PCR. Data are the mean ± standard deviation. Statistical significance was analyzed by analysis of variance. * p
    Figure Legend Snippet: Overexpression and knockdown of GLUT3 regulates stemness markers in CRC cells. ( A ) HCT116 cells were transfected with GLUT3/pEJ-3HA (2 µg) for 24 h. Western blot analysis was conducted for measuring the protein levels of GLUT3, Nanog and OCT3/4. ( B ) qRT-PCR was conducted to measure the mRNA levels of GLUT3, Bmi1, Nanog and OCT3/4. ( C ) Western blot analysis was conducted for measuring the protein levels of GLUT3, Nanog and OCT3/4 in shGLUT3-transfected SW620 cells. ( D ) qRT-PCR was conducted to measure the mRNA levels of GLUT3, Bmi1, Nanog and OCT3/4 in shGLUT3-transfected SW620 cells. GLUT3, glucose transporter 3; CRC, colorectal cancer; qRT-PCR, quantitative reverse transcription PCR. Data are the mean ± standard deviation. Statistical significance was analyzed by analysis of variance. * p

    Techniques Used: Over Expression, Transfection, Western Blot, Quantitative RT-PCR, Polymerase Chain Reaction, Standard Deviation

    ATF2 directly regulates GLUT3 transcription through TGF-β activation. ( A ) Lysates of the HCT116 cells treated or not with 10 μM SB505124 for 48 h. Western blot analysis was conducted for measuring the protein levels of TGF-β RI, GLUT3, p-ATF2, ATF2, p-JNK and JNK. ( B ) qRT-PCR was conducted to measure the mRNA levels of GLUT3 after being treated or not with TGF-β (10 ng/mL) for 24 h. ( C ) HCT116 cells were transiently transfected with GLUT3/pGL3 and ATF/CRE mutant GLUT3/pGL3 luciferase promoter plasmids for 24 h and treated or not with TGF-β (10 ng/mL) for 24 h. Cell extracts were harvested, and the luciferase assay was performed. ( D ) Effect of TGF-β activation on ATF2 binding to the GLUT3 chromatin. Cross-linked chromatin was immunoprecipitated with antibodies against ATF2 or rabbit lgG and analyzed by RT-PCR using primers that flank the ATF/CRE binding site. ATF2, activating transcription factor-2; p-ATF2, phospho-ATF2; GLUT3, glucose transporter 3; TGF-β, transforming growth factor-β; TGF-β RI, transforming growth factor-β receptor I; JNK, c-Jun N-terminal kinase; p-JNK, phospho-JNK; CRE, cAMP-response element; qRT-PCR, quantitative reverse transcription PCR; IgG, immunoglobulin G. Data are the mean ± standard deviation. Statistical significance was analyzed by analysis of variance. * p
    Figure Legend Snippet: ATF2 directly regulates GLUT3 transcription through TGF-β activation. ( A ) Lysates of the HCT116 cells treated or not with 10 μM SB505124 for 48 h. Western blot analysis was conducted for measuring the protein levels of TGF-β RI, GLUT3, p-ATF2, ATF2, p-JNK and JNK. ( B ) qRT-PCR was conducted to measure the mRNA levels of GLUT3 after being treated or not with TGF-β (10 ng/mL) for 24 h. ( C ) HCT116 cells were transiently transfected with GLUT3/pGL3 and ATF/CRE mutant GLUT3/pGL3 luciferase promoter plasmids for 24 h and treated or not with TGF-β (10 ng/mL) for 24 h. Cell extracts were harvested, and the luciferase assay was performed. ( D ) Effect of TGF-β activation on ATF2 binding to the GLUT3 chromatin. Cross-linked chromatin was immunoprecipitated with antibodies against ATF2 or rabbit lgG and analyzed by RT-PCR using primers that flank the ATF/CRE binding site. ATF2, activating transcription factor-2; p-ATF2, phospho-ATF2; GLUT3, glucose transporter 3; TGF-β, transforming growth factor-β; TGF-β RI, transforming growth factor-β receptor I; JNK, c-Jun N-terminal kinase; p-JNK, phospho-JNK; CRE, cAMP-response element; qRT-PCR, quantitative reverse transcription PCR; IgG, immunoglobulin G. Data are the mean ± standard deviation. Statistical significance was analyzed by analysis of variance. * p

    Techniques Used: Activation Assay, Western Blot, Quantitative RT-PCR, Transfection, Mutagenesis, Luciferase, Binding Assay, Immunoprecipitation, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Standard Deviation

    JNK is an upstream kinase regulating ATF2 in GLUT3-induced EMT in CRC cells. ( A ) Lysates of the HCT116 cells treated or not with SP600125 (20 μM) for 24 h. Western blot analysis was conducted for measuring the protein levels of p-JNK, JNK, p-ATF2, ATF2 and GLUT3. ( B ) Cell invasion was analyzed after 24 h post transfection with GLUT3/pEJ-3HA (2 µg) and treated or not with SP600125 (20 μM) for 24 h to allow for the permeabilization of the transwell membrane. The membrane was stained with 0.2% crystal violet. Scale bar, 100 µm. JNK, c-Jun N-terminal kinase; p-JNK, phospho-JNK; ATF2, activating transcription factor-2; p-ATF2, phospho-ATF2; GLUT3, glucose transporter 3; CRC, colorectal cancer. Data are the mean ± standard deviation. Statistical significance was analyzed by analysis of variance. ** p
    Figure Legend Snippet: JNK is an upstream kinase regulating ATF2 in GLUT3-induced EMT in CRC cells. ( A ) Lysates of the HCT116 cells treated or not with SP600125 (20 μM) for 24 h. Western blot analysis was conducted for measuring the protein levels of p-JNK, JNK, p-ATF2, ATF2 and GLUT3. ( B ) Cell invasion was analyzed after 24 h post transfection with GLUT3/pEJ-3HA (2 µg) and treated or not with SP600125 (20 μM) for 24 h to allow for the permeabilization of the transwell membrane. The membrane was stained with 0.2% crystal violet. Scale bar, 100 µm. JNK, c-Jun N-terminal kinase; p-JNK, phospho-JNK; ATF2, activating transcription factor-2; p-ATF2, phospho-ATF2; GLUT3, glucose transporter 3; CRC, colorectal cancer. Data are the mean ± standard deviation. Statistical significance was analyzed by analysis of variance. ** p

    Techniques Used: Western Blot, Transfection, Staining, Standard Deviation

    GLUT3 promotes EMT in HCT116 cells. ( A ) HCT116 cells were transfected with GLUT3/pEJ-3HA (2 µg) for 24 h. Western blot analysis was performed to measure the protein levels of GLUT3, E-cadherin, N-cadherin, snail, twist, α-SMA, CTGF, fibronectin, vimentin, PAI-1 and ZEB1. ( B ) qRT-PCR was performed to measure the mRNA levels of GLUT3, E-cadherin (E-cad), N-cadherin (N-Cad), snail, twist, α-SMA, CTGF, fibronectin 1 (FN1) and vimentin (Vim), PAI-1 and ZEB1. ( C ) The E-cad-Luc or α-SMA-Luc plasmids were transfected with GLUT3/pEJ-3HA into HCT116 cells for 24 h. Cell extracts were harvested, and the luciferase assay was performed. ( D ) Cell invasion was analyzed after 48 h post transfection with GLUT3/pEJ-3HA (2 µg) to allow for the permeabilization of the transwell membrane. The membrane was stained with 0.2% crystal violet. Scale bar, 100 µm. GLUT3, glucose transporter 3; α-SMA, α-smooth muscle actin; PAI-1, plasminogen activator inhibitor-1; ZEB1, zinc finger E-box binding homeobox 1, qRT-PCR, quantitative reverse transcription PCR; CTGF, connective tissue growth factor. Data are the mean ± standard deviation. Statistical significance was analyzed by analysis of variance. * p
    Figure Legend Snippet: GLUT3 promotes EMT in HCT116 cells. ( A ) HCT116 cells were transfected with GLUT3/pEJ-3HA (2 µg) for 24 h. Western blot analysis was performed to measure the protein levels of GLUT3, E-cadherin, N-cadherin, snail, twist, α-SMA, CTGF, fibronectin, vimentin, PAI-1 and ZEB1. ( B ) qRT-PCR was performed to measure the mRNA levels of GLUT3, E-cadherin (E-cad), N-cadherin (N-Cad), snail, twist, α-SMA, CTGF, fibronectin 1 (FN1) and vimentin (Vim), PAI-1 and ZEB1. ( C ) The E-cad-Luc or α-SMA-Luc plasmids were transfected with GLUT3/pEJ-3HA into HCT116 cells for 24 h. Cell extracts were harvested, and the luciferase assay was performed. ( D ) Cell invasion was analyzed after 48 h post transfection with GLUT3/pEJ-3HA (2 µg) to allow for the permeabilization of the transwell membrane. The membrane was stained with 0.2% crystal violet. Scale bar, 100 µm. GLUT3, glucose transporter 3; α-SMA, α-smooth muscle actin; PAI-1, plasminogen activator inhibitor-1; ZEB1, zinc finger E-box binding homeobox 1, qRT-PCR, quantitative reverse transcription PCR; CTGF, connective tissue growth factor. Data are the mean ± standard deviation. Statistical significance was analyzed by analysis of variance. * p

    Techniques Used: Transfection, Western Blot, Quantitative RT-PCR, Luciferase, Staining, Binding Assay, Polymerase Chain Reaction, Standard Deviation

    Knockdown of GLUT3 in SW620 cells reduces EMT process. ( A ) Western blot analysis was conducted for measuring the protein levels of GLUT3, E-cadherin, N-cadherin, snail, twist, α-SMA, CTGF, fibronectin 1 and vimentin, PAI-1 and ZEB1 in shGLUT3-induced SW620 cells. ( B ) qRT-PCR was conducted to measure the mRNA levels of GLUT3, E-cadherin (E-cad), N-cadherin (N-Cad), snail, twist, α-SMA, CTGF, fibronectin 1 (FN1) and vimentin (Vim), PAI-1 and ZEB1 in shGLUT3-transfected SW620 cells. ( C ) shGLUT3 was analyzed after 48 h using cell invasion assay to allow for the permeabilization of the transwell membrane. The membrane was stained with 0.2% crystal violet. Scale bar, 100 µm. GLUT3, glucose transporter 3; EMT, epithelial–mesenchymal transition; α-SMA, α-smooth muscle actin; PAI-1, plasminogen activator inhibitor-1; ZEB1, zinc finger E-box binding homeobox 1, qRT-PCR, quantitative reverse transcription PCR; CTGF, connective tissue growth factor. Data are the mean ± standard deviation. Statistical significance was analyzed by analysis of variance. * p
    Figure Legend Snippet: Knockdown of GLUT3 in SW620 cells reduces EMT process. ( A ) Western blot analysis was conducted for measuring the protein levels of GLUT3, E-cadherin, N-cadherin, snail, twist, α-SMA, CTGF, fibronectin 1 and vimentin, PAI-1 and ZEB1 in shGLUT3-induced SW620 cells. ( B ) qRT-PCR was conducted to measure the mRNA levels of GLUT3, E-cadherin (E-cad), N-cadherin (N-Cad), snail, twist, α-SMA, CTGF, fibronectin 1 (FN1) and vimentin (Vim), PAI-1 and ZEB1 in shGLUT3-transfected SW620 cells. ( C ) shGLUT3 was analyzed after 48 h using cell invasion assay to allow for the permeabilization of the transwell membrane. The membrane was stained with 0.2% crystal violet. Scale bar, 100 µm. GLUT3, glucose transporter 3; EMT, epithelial–mesenchymal transition; α-SMA, α-smooth muscle actin; PAI-1, plasminogen activator inhibitor-1; ZEB1, zinc finger E-box binding homeobox 1, qRT-PCR, quantitative reverse transcription PCR; CTGF, connective tissue growth factor. Data are the mean ± standard deviation. Statistical significance was analyzed by analysis of variance. * p

    Techniques Used: Western Blot, Quantitative RT-PCR, Transfection, Invasion Assay, Staining, Binding Assay, Polymerase Chain Reaction, Standard Deviation

    TGF-β signaling pathway regulates GLUT3-induced EMT in CRC cells. ( A ) Lysates of the HCT116 cells treated or not with 10 μM SB431542 for 48 h. Western blot analysis was conducted for measuring the protein levels of TGF-β RI, GLUT3, α-SMA, ZEB1 and PAI-1. ( B ) Cell invasion was analyzed after 24 h post transfection with GLUT3/pEJ-3HA (1.5 µg) and treated or not with 10 μM SB431542 for 24 h to allow for the permeabilization of the transwell membrane. The membrane was stained with 0.2% crystal violet. Scale bar, 100 µm. TGF-β, transforming growth factor-β; GLUT3, glucose transporter 3; EMT, epithelial–mesenchymal transition; TGF-β RI, transforming growth factor-β receptor I; α-SMA, α-smooth muscle actin; ZEB1, zinc finger E-box binding homeobox 1, PAI-1, plasminogen activator inhibitor-1. Data are the mean ± standard deviation. Statistical significance was analyzed by analysis of variance. * p
    Figure Legend Snippet: TGF-β signaling pathway regulates GLUT3-induced EMT in CRC cells. ( A ) Lysates of the HCT116 cells treated or not with 10 μM SB431542 for 48 h. Western blot analysis was conducted for measuring the protein levels of TGF-β RI, GLUT3, α-SMA, ZEB1 and PAI-1. ( B ) Cell invasion was analyzed after 24 h post transfection with GLUT3/pEJ-3HA (1.5 µg) and treated or not with 10 μM SB431542 for 24 h to allow for the permeabilization of the transwell membrane. The membrane was stained with 0.2% crystal violet. Scale bar, 100 µm. TGF-β, transforming growth factor-β; GLUT3, glucose transporter 3; EMT, epithelial–mesenchymal transition; TGF-β RI, transforming growth factor-β receptor I; α-SMA, α-smooth muscle actin; ZEB1, zinc finger E-box binding homeobox 1, PAI-1, plasminogen activator inhibitor-1. Data are the mean ± standard deviation. Statistical significance was analyzed by analysis of variance. * p

    Techniques Used: Western Blot, Transfection, Staining, Binding Assay, Standard Deviation

    7) Product Images from "Myricetin induces apoptosis and autophagy by inhibiting PI3K/Akt/mTOR signalling in human colon cancer cells"

    Article Title: Myricetin induces apoptosis and autophagy by inhibiting PI3K/Akt/mTOR signalling in human colon cancer cells

    Journal: BMC Complementary Medicine and Therapies

    doi: 10.1186/s12906-020-02965-w

    Myricetin inhibited the viability of human colorectal cancer cells. HCT116 and SW620 cells were treated with 0–400 μmol/L myricetin for 24 h, 48 h, and 72 h. Cell viability was analysed by means of a resazurin cell viability assay. Three replicate wells were set up in 96-well plates for each experimental group, and the experiment was repeated three times
    Figure Legend Snippet: Myricetin inhibited the viability of human colorectal cancer cells. HCT116 and SW620 cells were treated with 0–400 μmol/L myricetin for 24 h, 48 h, and 72 h. Cell viability was analysed by means of a resazurin cell viability assay. Three replicate wells were set up in 96-well plates for each experimental group, and the experiment was repeated three times

    Techniques Used: Viability Assay

    8) Product Images from "miR-145 Antagonizes SNAI1-Mediated Stemness and Radiation Resistance in Colorectal Cancer"

    Article Title: miR-145 Antagonizes SNAI1-Mediated Stemness and Radiation Resistance in Colorectal Cancer

    Journal: Molecular Therapy

    doi: 10.1016/j.ymthe.2017.12.023

    SNAI1 Represses miR-145 Promoter Activity and Expression in Colorectal Cancer Cells
    Figure Legend Snippet: SNAI1 Represses miR-145 Promoter Activity and Expression in Colorectal Cancer Cells

    Techniques Used: Activity Assay, Expressing

    Detection of Functional SNAI1:miR145 Axis in EpCAM+/ALDH+ Enriched CSC Population in Colorectal Cancer PDX Tumors
    Figure Legend Snippet: Detection of Functional SNAI1:miR145 Axis in EpCAM+/ALDH+ Enriched CSC Population in Colorectal Cancer PDX Tumors

    Techniques Used: Functional Assay

    Detection of Functional SNAI1:miR145 Axis in EpCAM+/ALDH+ Enriched CSC Population in Colorectal Cancer PDX Tumors
    Figure Legend Snippet: Detection of Functional SNAI1:miR145 Axis in EpCAM+/ALDH+ Enriched CSC Population in Colorectal Cancer PDX Tumors

    Techniques Used: Functional Assay

    Detection of Functional SNAI1:miR145 Axis in EpCAM+/ALDH+ Enriched CSC Population in Colorectal Cancer PDX Tumors
    Figure Legend Snippet: Detection of Functional SNAI1:miR145 Axis in EpCAM+/ALDH+ Enriched CSC Population in Colorectal Cancer PDX Tumors

    Techniques Used: Functional Assay

    Therapeutic Mechanism of miR-145 Delivery to Overcome SNAI1-Mediated Colorectal Cancer Radiation Resistance
    Figure Legend Snippet: Therapeutic Mechanism of miR-145 Delivery to Overcome SNAI1-Mediated Colorectal Cancer Radiation Resistance

    Techniques Used:

    Elevated SNAI1 Expression in Human Colorectal Cancer Datasets
    Figure Legend Snippet: Elevated SNAI1 Expression in Human Colorectal Cancer Datasets

    Techniques Used: Expressing

    miR-145 Sensitizes SNAI1-Overexpressing Colorectal Cancer Cells to Radiation Therapy
    Figure Legend Snippet: miR-145 Sensitizes SNAI1-Overexpressing Colorectal Cancer Cells to Radiation Therapy

    Techniques Used:

    9) Product Images from "miR-191 promotes tumorigenesis of human colorectal cancer through targeting C/EBPβ"

    Article Title: miR-191 promotes tumorigenesis of human colorectal cancer through targeting C/EBPβ

    Journal: Oncotarget

    doi:

    Schematic model of miR-191-mediated promotion of tumorigencity of colorectal cancer cells In colorectal cancer cells, various anticancer drugs regulate the expression of miR-191. miR-191 causes a number of genes to respond and adapt. On one hand, miR-191 induces the expression of CDK4 to increase cell growth. On the other hand, miR-191 suppresses the level of C/EBPβ, a tumor suppressor gene functions as a transcriptional activator of p15, p16 and p57, which are the key regulators of cell cycle and cell survival. As a result, miR-191 induces the cell cycle progression and tumor cell resistance to various stimuli.
    Figure Legend Snippet: Schematic model of miR-191-mediated promotion of tumorigencity of colorectal cancer cells In colorectal cancer cells, various anticancer drugs regulate the expression of miR-191. miR-191 causes a number of genes to respond and adapt. On one hand, miR-191 induces the expression of CDK4 to increase cell growth. On the other hand, miR-191 suppresses the level of C/EBPβ, a tumor suppressor gene functions as a transcriptional activator of p15, p16 and p57, which are the key regulators of cell cycle and cell survival. As a result, miR-191 induces the cell cycle progression and tumor cell resistance to various stimuli.

    Techniques Used: Expressing

    miR-191 is up-regulated in colon cancers Stem-loop RT-PCR analysis of the miR-191 level in tissues and cell lines. (A) Relative miR-191 expression in 16 paired colon cancer tissues and adjacent non-tumor tissues (upper panel). miR-191 expression values were expressed as ratios with U6 snRNA (×10). (lower panel, P = 0.011) The statistical significance was evaluated by paired-samples T test. (B) Relative miR-191 expression in five human colorectal cancer cell lines (HCT116, RKO, HT29, SW480, DLD1 and Lovo) and human embryonic kidney 293T cells. U6 snRNA was used as an internal control. The data represents the means ± SDs.
    Figure Legend Snippet: miR-191 is up-regulated in colon cancers Stem-loop RT-PCR analysis of the miR-191 level in tissues and cell lines. (A) Relative miR-191 expression in 16 paired colon cancer tissues and adjacent non-tumor tissues (upper panel). miR-191 expression values were expressed as ratios with U6 snRNA (×10). (lower panel, P = 0.011) The statistical significance was evaluated by paired-samples T test. (B) Relative miR-191 expression in five human colorectal cancer cell lines (HCT116, RKO, HT29, SW480, DLD1 and Lovo) and human embryonic kidney 293T cells. U6 snRNA was used as an internal control. The data represents the means ± SDs.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing

    10) Product Images from "Myricetin induces apoptosis and autophagy by inhibiting PI3K/Akt/mTOR signalling in human colon cancer cells"

    Article Title: Myricetin induces apoptosis and autophagy by inhibiting PI3K/Akt/mTOR signalling in human colon cancer cells

    Journal: BMC Complementary Medicine and Therapies

    doi: 10.1186/s12906-020-02965-w

    Myricetin inhibited the viability of human colorectal cancer cells. HCT116 and SW620 cells were treated with 0–400 μmol/L myricetin for 24 h, 48 h, and 72 h. Cell viability was analysed by means of a resazurin cell viability assay. Three replicate wells were set up in 96-well plates for each experimental group, and the experiment was repeated three times
    Figure Legend Snippet: Myricetin inhibited the viability of human colorectal cancer cells. HCT116 and SW620 cells were treated with 0–400 μmol/L myricetin for 24 h, 48 h, and 72 h. Cell viability was analysed by means of a resazurin cell viability assay. Three replicate wells were set up in 96-well plates for each experimental group, and the experiment was repeated three times

    Techniques Used: Viability Assay

    11) Product Images from "MicroRNA-375 suppresses human colorectal cancer metastasis by targeting Frizzled 8"

    Article Title: MicroRNA-375 suppresses human colorectal cancer metastasis by targeting Frizzled 8

    Journal: Oncotarget

    doi: 10.18632/oncotarget.9811

    Low miR-375 expression predicts metastatic potential in human colorectal cancer (CRC) patients A. miR-375 expression was measured in 90 paired human CRC and adjacent noncancerous tissues (NCTs) by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Two–tailed non-parametric Wilcoxon test were used to evaluate the statistical differences of miR-375 levels. Specifically, miR-375 expression was markedly downregulated in cancer tissues compared with their corresponding NCTs. U6 small nuclear RNA was used an as internal control (**** p
    Figure Legend Snippet: Low miR-375 expression predicts metastatic potential in human colorectal cancer (CRC) patients A. miR-375 expression was measured in 90 paired human CRC and adjacent noncancerous tissues (NCTs) by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Two–tailed non-parametric Wilcoxon test were used to evaluate the statistical differences of miR-375 levels. Specifically, miR-375 expression was markedly downregulated in cancer tissues compared with their corresponding NCTs. U6 small nuclear RNA was used an as internal control (**** p

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Two Tailed Test

    12) Product Images from "Upregulated plasmacytoma variant translocation 1 promotes cell proliferation, invasion and metastasis in colorectal cancer"

    Article Title: Upregulated plasmacytoma variant translocation 1 promotes cell proliferation, invasion and metastasis in colorectal cancer

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2018.8669

    Knocking down PVT1 expression inhibits invasion of colorectal cancer cells. Cell invasive ability was determined by transwell Matrigel assay after transfecting with si-NC or si- PVT1 for 48 h in HCT116 cells. Data are shown as the mean ± standard error of the mean. **P
    Figure Legend Snippet: Knocking down PVT1 expression inhibits invasion of colorectal cancer cells. Cell invasive ability was determined by transwell Matrigel assay after transfecting with si-NC or si- PVT1 for 48 h in HCT116 cells. Data are shown as the mean ± standard error of the mean. **P

    Techniques Used: Expressing, Matrigel Assay

    Knocking down PVT1 expression inhibits proliferation of colorectal cancer cells. (A) Interference efficiency of si- PVT1 was verified in HCT116 cells. HCT116 cells were transfected with either si-NC or si- PVT1 (1 # , 2 # , 1+2 # ) for 48 h, and then PVT1 expression was analysed by reverse transcription-quantitative polymerase chain reaction. (B and C) CCK-8 assay and cell clone-formation assay was used to detect the cell proliferative ability after transfecting with si-NC or si- PVT1 for 48 h in HCT116 cells. Data are shown as the mean ± standard error of the mean. *P
    Figure Legend Snippet: Knocking down PVT1 expression inhibits proliferation of colorectal cancer cells. (A) Interference efficiency of si- PVT1 was verified in HCT116 cells. HCT116 cells were transfected with either si-NC or si- PVT1 (1 # , 2 # , 1+2 # ) for 48 h, and then PVT1 expression was analysed by reverse transcription-quantitative polymerase chain reaction. (B and C) CCK-8 assay and cell clone-formation assay was used to detect the cell proliferative ability after transfecting with si-NC or si- PVT1 for 48 h in HCT116 cells. Data are shown as the mean ± standard error of the mean. *P

    Techniques Used: Expressing, Transfection, Real-time Polymerase Chain Reaction, CCK-8 Assay, Tube Formation Assay

    13) Product Images from "NQO1 inhibits proteasome-mediated degradation of HIF-1α"

    Article Title: NQO1 inhibits proteasome-mediated degradation of HIF-1α

    Journal: Nature Communications

    doi: 10.1038/ncomms13593

    Upregulation of NQO1 correlates with poor prognosis and expression of HIF-1α in colorectal cancer. ( a ) An Oncomine analysis of the TCGA colorectal database indicated that NQO1 expressions are elevated in colorectal cancers ( n =102) compared with normal colorectal tissues ( n =19). **** P
    Figure Legend Snippet: Upregulation of NQO1 correlates with poor prognosis and expression of HIF-1α in colorectal cancer. ( a ) An Oncomine analysis of the TCGA colorectal database indicated that NQO1 expressions are elevated in colorectal cancers ( n =102) compared with normal colorectal tissues ( n =19). **** P

    Techniques Used: Expressing

    14) Product Images from "pH-responsive Virus-like Nanoparticles with Enhanced Tumour-targeting Ligands for Cancer Drug Delivery"

    Article Title: pH-responsive Virus-like Nanoparticles with Enhanced Tumour-targeting Ligands for Cancer Drug Delivery

    Journal: Scientific Reports

    doi: 10.1038/srep37891

    Live cell imaging analysis of the delivery of doxorubicin by tHBcAg nanoparticles into colorectal cancer and normal cells. ( a ) Colorectal cancer HT29 and ( b ) Caco-2 cells and ( c ) normal CCD-112 cells were incubated with free doxorubicin (DOX), tHBcAg nanoparticles loaded with PAA-DOX (tHBcAg-PAA-DOX), folic acid (FA)-conjugated tHBcAg nanoparticles loaded with PAA-DOX (FA-tHBcAg-PAA-DOX), and FA-conjugated tHBcAg nanoparticles using the nanoglue and loaded with PAA-DOX (FA-N-tHBcAg-PAA-DOX) at equivalent DOX concentrations (5 μg/mL) at 37 °C and 5% CO 2 for 1 h. The untreated cells were used as negative controls. The samples are labelled on the left. Nuclei were stained with Hoechst (blue), and DOX was excited at 480 nm. Scale bars indicate 20 μm.
    Figure Legend Snippet: Live cell imaging analysis of the delivery of doxorubicin by tHBcAg nanoparticles into colorectal cancer and normal cells. ( a ) Colorectal cancer HT29 and ( b ) Caco-2 cells and ( c ) normal CCD-112 cells were incubated with free doxorubicin (DOX), tHBcAg nanoparticles loaded with PAA-DOX (tHBcAg-PAA-DOX), folic acid (FA)-conjugated tHBcAg nanoparticles loaded with PAA-DOX (FA-tHBcAg-PAA-DOX), and FA-conjugated tHBcAg nanoparticles using the nanoglue and loaded with PAA-DOX (FA-N-tHBcAg-PAA-DOX) at equivalent DOX concentrations (5 μg/mL) at 37 °C and 5% CO 2 for 1 h. The untreated cells were used as negative controls. The samples are labelled on the left. Nuclei were stained with Hoechst (blue), and DOX was excited at 480 nm. Scale bars indicate 20 μm.

    Techniques Used: Live Cell Imaging, Incubation, Staining

    15) Product Images from "miR-1285-3p Controls Colorectal Cancer Proliferation and Escape from Apoptosis through DAPK2"

    Article Title: miR-1285-3p Controls Colorectal Cancer Proliferation and Escape from Apoptosis through DAPK2

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21072423

    MiR-1285 blockade by a lentiviral sponge construct phenocopies the anti-survival effect of LNA-mediated inhibition. ( a ) Schematic representation of the doxycycline-inducible expression construct for the miR-1285 sponge (Tet-ON system): TRE = tetracycline response element, rtTA3 = reverse tetracycline transactivator 3. The transactivator binds to the TRE in the presence of doxycycline. ( b ) FACS analysis of GFP expression upon doxycycline treatment of CR-CSC 2 cells transduced with control virus or anti-miR-1285 sponge. ( c ) Soft agar colony formation assays in both cancer stem cells and SW480. Cells were transduced with the indicated lentiviral constructs and treated with doxycycline. Mean values of one or two independent experiments performed in triplicate are shown for CR-CSCs and SW480 cells, respectively. The graph shows the percentage of plated cells that gave rise to colonies (mean ± SEM). ( d ) Assessment of cell number by Cell Titer Glo assay in transduced primary cancer stem cells (CR-CSC 2) following doxycycline administration. Mean values of two independent experiments performed in quadruplicate are shown (mean ± SEM). ( e ) Detection of PARP-1 cleavage by Western blot in transduced SW480 cells treated with doxycycline. The (+) and (-) symbols indicate the presence or absence of doxycycline. p -values were calculated by Student’s t test ( ns = not significant, ** p
    Figure Legend Snippet: MiR-1285 blockade by a lentiviral sponge construct phenocopies the anti-survival effect of LNA-mediated inhibition. ( a ) Schematic representation of the doxycycline-inducible expression construct for the miR-1285 sponge (Tet-ON system): TRE = tetracycline response element, rtTA3 = reverse tetracycline transactivator 3. The transactivator binds to the TRE in the presence of doxycycline. ( b ) FACS analysis of GFP expression upon doxycycline treatment of CR-CSC 2 cells transduced with control virus or anti-miR-1285 sponge. ( c ) Soft agar colony formation assays in both cancer stem cells and SW480. Cells were transduced with the indicated lentiviral constructs and treated with doxycycline. Mean values of one or two independent experiments performed in triplicate are shown for CR-CSCs and SW480 cells, respectively. The graph shows the percentage of plated cells that gave rise to colonies (mean ± SEM). ( d ) Assessment of cell number by Cell Titer Glo assay in transduced primary cancer stem cells (CR-CSC 2) following doxycycline administration. Mean values of two independent experiments performed in quadruplicate are shown (mean ± SEM). ( e ) Detection of PARP-1 cleavage by Western blot in transduced SW480 cells treated with doxycycline. The (+) and (-) symbols indicate the presence or absence of doxycycline. p -values were calculated by Student’s t test ( ns = not significant, ** p

    Techniques Used: Construct, Inhibition, Expressing, FACS, Transduction, Glo Assay, Western Blot

    DAPK2 over-expression recapitulates both the apoptosis induction and the cell cycle arrest observed upon miR-1285 inhibition. SW480 cells were transduced with a lentiviral construct for ectopic expression of DAPK2 and compared to their counterparts either harboring the empty vector (CTR) or untreated (NT). ( a ) Expression levels of DAPK2, PARP-1 and cleaved Caspase-3 in the indicated samples. GAPDH levels were assessed as a loading control. ( b ) Soft agar colony formation assay. The graph shows the percentage of plated cells that gave rise to colonies. ( c ) Cell cycle analysis showing cell populations in sub-G0, G1, S and G2/M phases: left panel shows one representative experiment, whereas the histograms show the mean values of four independent experiments. Error bars represent SEM values and p -values were calculated by Student’s t test (* p
    Figure Legend Snippet: DAPK2 over-expression recapitulates both the apoptosis induction and the cell cycle arrest observed upon miR-1285 inhibition. SW480 cells were transduced with a lentiviral construct for ectopic expression of DAPK2 and compared to their counterparts either harboring the empty vector (CTR) or untreated (NT). ( a ) Expression levels of DAPK2, PARP-1 and cleaved Caspase-3 in the indicated samples. GAPDH levels were assessed as a loading control. ( b ) Soft agar colony formation assay. The graph shows the percentage of plated cells that gave rise to colonies. ( c ) Cell cycle analysis showing cell populations in sub-G0, G1, S and G2/M phases: left panel shows one representative experiment, whereas the histograms show the mean values of four independent experiments. Error bars represent SEM values and p -values were calculated by Student’s t test (* p

    Techniques Used: Over Expression, Inhibition, Transduction, Construct, Expressing, Plasmid Preparation, Soft Agar Assay, Cell Cycle Assay

    DAPK2 is a direct target of miR-1285 and mediates apoptosis induction in miR-1285 depleted cells. ( a ) Western blot showing an increase in DAPK2 levels and concomitant PARP-1 cleavage in SW480 cells upon transfection with LNA-1285 at 25 nM for 24 h and in untreated cells (-). The same protein samples were loaded in two different gels. GAPDH levels are shown as a loading control. The densitometric analysis is reported as fold change compared to the sample treated with control LNA. ( b ) RT-qPCR for DAPK2 in SW480 cells untreated (-) or transfected with the indicated LNA at 25 nM for 72 h. ( c ) Western blot showing the increase in DAPK2 levels in primary CR-CSC 2 transduced with the anti-miR-1285 lentiviral sponge construct, control construct or untreated (-), upon doxycycline treatment. ( d ) Luciferase activity in HeLa cells transfected with DAPK2 3′UTR in combination with control or anti-miR-1285 LNA. The graph shows the normalized luciferase activity of three independent experiments performed in quadruplicate (mean ± SEM). ( e , f ) Cell Titer-Glo assay and Western blot show that down-modulation of DAPK2 by RNA interference can rescue the LNA-1285 apoptotic effect in SW480 cells. The (-) symbol indicates untreated cells. ( e ) Cell number was measured 72 h upon transfection with the indicated molecules (mean ± SEM). ( f ) Western blotting showing DAPK2 modulation in SW480 cells after the indicated treatments. Double transfection of anti-DAPK2 siRNA together with LNA-1285 almost restores the DAPK2 levels of the control cells, counteracting LNA-mediated induction. CASP-3 cleavage is reduced to ~60%. TUBULIN was used as the protein-loading control. Numbers indicate DAPK2 levels as the fold change relative to the control-treated cells, and cleaved CASP-3 levels as the fold decrease relative to the LNA-1285/control siRNA-treated cells. The values were normalized to the TUBULIN signal, as measured by densitometric analyses. The second gel image was cropped in order to display the samples in an order different from the one of the loading. Tubulin levels are shown as a loading control. p -values were calculated by Student’s t test (** p
    Figure Legend Snippet: DAPK2 is a direct target of miR-1285 and mediates apoptosis induction in miR-1285 depleted cells. ( a ) Western blot showing an increase in DAPK2 levels and concomitant PARP-1 cleavage in SW480 cells upon transfection with LNA-1285 at 25 nM for 24 h and in untreated cells (-). The same protein samples were loaded in two different gels. GAPDH levels are shown as a loading control. The densitometric analysis is reported as fold change compared to the sample treated with control LNA. ( b ) RT-qPCR for DAPK2 in SW480 cells untreated (-) or transfected with the indicated LNA at 25 nM for 72 h. ( c ) Western blot showing the increase in DAPK2 levels in primary CR-CSC 2 transduced with the anti-miR-1285 lentiviral sponge construct, control construct or untreated (-), upon doxycycline treatment. ( d ) Luciferase activity in HeLa cells transfected with DAPK2 3′UTR in combination with control or anti-miR-1285 LNA. The graph shows the normalized luciferase activity of three independent experiments performed in quadruplicate (mean ± SEM). ( e , f ) Cell Titer-Glo assay and Western blot show that down-modulation of DAPK2 by RNA interference can rescue the LNA-1285 apoptotic effect in SW480 cells. The (-) symbol indicates untreated cells. ( e ) Cell number was measured 72 h upon transfection with the indicated molecules (mean ± SEM). ( f ) Western blotting showing DAPK2 modulation in SW480 cells after the indicated treatments. Double transfection of anti-DAPK2 siRNA together with LNA-1285 almost restores the DAPK2 levels of the control cells, counteracting LNA-mediated induction. CASP-3 cleavage is reduced to ~60%. TUBULIN was used as the protein-loading control. Numbers indicate DAPK2 levels as the fold change relative to the control-treated cells, and cleaved CASP-3 levels as the fold decrease relative to the LNA-1285/control siRNA-treated cells. The values were normalized to the TUBULIN signal, as measured by densitometric analyses. The second gel image was cropped in order to display the samples in an order different from the one of the loading. Tubulin levels are shown as a loading control. p -values were calculated by Student’s t test (** p

    Techniques Used: Western Blot, Transfection, Quantitative RT-PCR, Transduction, Construct, Luciferase, Activity Assay, Glo Assay

    Transcriptome analyses upon LNA-1285 treatment. ( a ) Heat map showing unsupervised hierarchical clustering of all modulated genes in LNA-1285-treated SW480 cells compared to control cells (untreated + control LNA). ( b ) Volcano plot representing all modulated genes (upregulated in red and downregulated in green). ( c ) Gene set enrichment analysis (GSEA) for differential gene expression in miR-1285-depleted cells shows significant modulation of genes related to apoptosis and cell cycle arrest. FDR q-val
    Figure Legend Snippet: Transcriptome analyses upon LNA-1285 treatment. ( a ) Heat map showing unsupervised hierarchical clustering of all modulated genes in LNA-1285-treated SW480 cells compared to control cells (untreated + control LNA). ( b ) Volcano plot representing all modulated genes (upregulated in red and downregulated in green). ( c ) Gene set enrichment analysis (GSEA) for differential gene expression in miR-1285-depleted cells shows significant modulation of genes related to apoptosis and cell cycle arrest. FDR q-val

    Techniques Used: Expressing

    MiR-1285 inhibition impairs cell survival/proliferation and clonogenic potential in colorectal cancer cell lines and patient-derived cells. ( a ) Diagram showing results of the functional anti-miR screening in SW480 cells (906 LNAs tested). The viable cell number was evaluated 72 h after transfection by Cell Titer-Glo luminescent cell viability assay and normalized to the average of all samples, excluding controls. Each dot represents a single LNA. Values above and below two-fold the standard deviation (red dashed lines) were considered as significant. The experiment was performed in triplicate and the mean values ± SD were plotted. ( b ) Validation of the screening results. Cell number was evaluated by Cell Titer Glo assay 72 h after transfection of SW480 cells with the indicated LNAs at 10 nM and percentages versus the untreated sample (-) are reported (mean ± SEM). ( c ) Growth curves of both SW480 and colorectal cancer stem cells (CR-CSC 1) untreated (nt) and following treatment with LNA-1285 or control LNA at 5 nM and 25 nM, respectively. Statistical analysis was performed by ANOVA test for unpaired groups (** p
    Figure Legend Snippet: MiR-1285 inhibition impairs cell survival/proliferation and clonogenic potential in colorectal cancer cell lines and patient-derived cells. ( a ) Diagram showing results of the functional anti-miR screening in SW480 cells (906 LNAs tested). The viable cell number was evaluated 72 h after transfection by Cell Titer-Glo luminescent cell viability assay and normalized to the average of all samples, excluding controls. Each dot represents a single LNA. Values above and below two-fold the standard deviation (red dashed lines) were considered as significant. The experiment was performed in triplicate and the mean values ± SD were plotted. ( b ) Validation of the screening results. Cell number was evaluated by Cell Titer Glo assay 72 h after transfection of SW480 cells with the indicated LNAs at 10 nM and percentages versus the untreated sample (-) are reported (mean ± SEM). ( c ) Growth curves of both SW480 and colorectal cancer stem cells (CR-CSC 1) untreated (nt) and following treatment with LNA-1285 or control LNA at 5 nM and 25 nM, respectively. Statistical analysis was performed by ANOVA test for unpaired groups (** p

    Techniques Used: Inhibition, Derivative Assay, Functional Assay, Transfection, Cell Viability Assay, Standard Deviation, Glo Assay

    Targeting miR-1285 by an LNA-based anti-miR induces apoptosis and cell cycle arrest. ( a ) PI/Annexin V staining in SW480 cells: the dot plot shows a representative experiment, whereas the mean values of two different experiments are plotted in the graph (mean ± SEM). ( b ) Total and cleaved Caspase-3 levels were assayed by Western blot; nucleolin was detected as a loading control. ( c ) PI/Annexin V staining in colorectal cancer stem cells. Mean values of three independent experiments are shown (mean ± SEM). ( d ) FACS analysis of cell cycle in SW480 cells untreated (-) and transfected with control LNA or LNA-1285. The graph shows the percentage of cells in the different phases of the cell cycle. Error bars represent SEM. ( e ) Western blot showing reduced levels of phospho-RB and Cyclin B1 upon miR-1285 depletion in CR-CSC 2. HSP90 levels are shown as loading control. The (-) symbol indicates the untreated control. p -values were calculated by Student’s t test (** p
    Figure Legend Snippet: Targeting miR-1285 by an LNA-based anti-miR induces apoptosis and cell cycle arrest. ( a ) PI/Annexin V staining in SW480 cells: the dot plot shows a representative experiment, whereas the mean values of two different experiments are plotted in the graph (mean ± SEM). ( b ) Total and cleaved Caspase-3 levels were assayed by Western blot; nucleolin was detected as a loading control. ( c ) PI/Annexin V staining in colorectal cancer stem cells. Mean values of three independent experiments are shown (mean ± SEM). ( d ) FACS analysis of cell cycle in SW480 cells untreated (-) and transfected with control LNA or LNA-1285. The graph shows the percentage of cells in the different phases of the cell cycle. Error bars represent SEM. ( e ) Western blot showing reduced levels of phospho-RB and Cyclin B1 upon miR-1285 depletion in CR-CSC 2. HSP90 levels are shown as loading control. The (-) symbol indicates the untreated control. p -values were calculated by Student’s t test (** p

    Techniques Used: Staining, Western Blot, FACS, Transfection

    16) Product Images from "The long noncoding RNA SNHG1 regulates colorectal cancer cell growth through interactions with EZH2 and miR-154-5p"

    Article Title: The long noncoding RNA SNHG1 regulates colorectal cancer cell growth through interactions with EZH2 and miR-154-5p

    Journal: Molecular Cancer

    doi: 10.1186/s12943-018-0894-x

    SNHG1 acts as a sponge for miR-154-5p in the cytoplasm. a Representative FISH images indicated subcellular location of SNHG1 in HCT-116 and HCT-8 cells (red). Nuclei were stained by DAPI (blue). SNHG1 sense probe was employed as a negative control. b Relative SNHG1 expression levels in nuclear and cytosolic fractions of HCT-116 and HCT-8 cells. Nuclear controls: U6; Cytosolic controls: GAPDH. c Dual luciferase reporter assays were used to determine miRNAs that directly interacted with SNHG1. Luciferase activity is presented as relative luciferase activity normalized to activity of their respective negative control. d Dual luciferase reporter assays were conducted with wild type and mutant type (putative binding sites for miR-154-5p were mutated) luciferase reporter vectors. Right panel, sequence alignment of miR-154-5p and its predicted binding sites (green) in SNHG1. Predicted miR-154-5p target sequence (blue) in SNHG1 (Luc-SNHG1-wt) and position of mutated nucleotides (red) in SNHG1 (Luc-SNHG1-mut). e RNA immunoprecipitation with an anti-Ago2 antibody was used to assess endogenous Ago2 binding to RNA in HCT-116 cells, IgG was used as the control. SNHG1 and miR-154-5p levels were determined by qRT–PCR and presented as fold enrichment in Ago2 relative to input. RIP efficiency of Ago2 protein was detected by western blot. f RNA pull-down assays were used to examine the interaction of SNHG1 and Ago2 in HCT-116 cells. g CCK-8 assays demonstrated that SNHG1 silencing inhibited HCT-116 cell growth. MiR-154-5p down-regulation rescued growth inhibition caused by SNHG1 knockdown. h EdU assays revealed that SNHG1 overexpression promotes HCT-116 cell proliferation. Co-transfecting miR-154-5p mimics with the SNHG1 plasmid abolished the increased proliferation rates. i The correlation between miR-154-5p and SNHG1 expression analyzed in 30 paired colorectal cancer samples (n = 30, r = − 0.48, P = 0.008). Scale bar = 50 μm. * P
    Figure Legend Snippet: SNHG1 acts as a sponge for miR-154-5p in the cytoplasm. a Representative FISH images indicated subcellular location of SNHG1 in HCT-116 and HCT-8 cells (red). Nuclei were stained by DAPI (blue). SNHG1 sense probe was employed as a negative control. b Relative SNHG1 expression levels in nuclear and cytosolic fractions of HCT-116 and HCT-8 cells. Nuclear controls: U6; Cytosolic controls: GAPDH. c Dual luciferase reporter assays were used to determine miRNAs that directly interacted with SNHG1. Luciferase activity is presented as relative luciferase activity normalized to activity of their respective negative control. d Dual luciferase reporter assays were conducted with wild type and mutant type (putative binding sites for miR-154-5p were mutated) luciferase reporter vectors. Right panel, sequence alignment of miR-154-5p and its predicted binding sites (green) in SNHG1. Predicted miR-154-5p target sequence (blue) in SNHG1 (Luc-SNHG1-wt) and position of mutated nucleotides (red) in SNHG1 (Luc-SNHG1-mut). e RNA immunoprecipitation with an anti-Ago2 antibody was used to assess endogenous Ago2 binding to RNA in HCT-116 cells, IgG was used as the control. SNHG1 and miR-154-5p levels were determined by qRT–PCR and presented as fold enrichment in Ago2 relative to input. RIP efficiency of Ago2 protein was detected by western blot. f RNA pull-down assays were used to examine the interaction of SNHG1 and Ago2 in HCT-116 cells. g CCK-8 assays demonstrated that SNHG1 silencing inhibited HCT-116 cell growth. MiR-154-5p down-regulation rescued growth inhibition caused by SNHG1 knockdown. h EdU assays revealed that SNHG1 overexpression promotes HCT-116 cell proliferation. Co-transfecting miR-154-5p mimics with the SNHG1 plasmid abolished the increased proliferation rates. i The correlation between miR-154-5p and SNHG1 expression analyzed in 30 paired colorectal cancer samples (n = 30, r = − 0.48, P = 0.008). Scale bar = 50 μm. * P

    Techniques Used: Fluorescence In Situ Hybridization, Staining, Negative Control, Expressing, Luciferase, Activity Assay, Mutagenesis, Binding Assay, Sequencing, Immunoprecipitation, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Inhibition, Over Expression, Plasmid Preparation

    SNHG 1 promotes colorectal cancer progression partly by regulating KLF2 and CDKN2B expression. a Left panel, CCK-8 assays demonstrated that silence of SNHG1 inhibited cancer cell growth. KLF2 knockdown could rescue growth inhibition caused by SNHG1 knockdown in HCT-116 cells. Right panel, CCK-8 assays demonstrated that silence of SNHG1 inhibited cancer cell grow th. CDKN2B (P15) knockdown could rescue growth inhibition caused by SNHG1 knockdown in HCT-116 cells. b EdU assays showed that SNHG1 knockdown inhibited cancer cell proliferation. Co-transfecting KLF2 or CDKN2B siRNAs with SNHG1 siRNAs reversed the decreased proliferation rates in HCT-116 cells. c EdU assays showed that EZH2 knockdown could inhibit proliferation promotion caused by SNHG1 overexpression in HCT-116 cells. d Immunohistochemistry analysis of EZH2, KLF2 and CDKN2B protein levels in colorectal cancer and normal tissues. e Immunohistochemistry analysis of KLF2 and CDKN2B protein levels in tumor tissues formed from SNHG1 knockdown or control cells. f Schematic of the proposed mechanism of SNHG1 in colorectal cancer cells. In the cytoplasm, SNHG1 acts as a ceRNA to sponge miR-154-5p and upregulated the expression of CCND2 (CyclinD2). In the nucleus, SNHG1 is involved in PRC2 mediated epigenetic repression of KLF2 and CDKN2B (P15). KLF2 is also an upstream regulatory factor of CDKN2B. Besides, CDKN2B is a well-studied inhibitor of CCND2. Downstream genes of SNHG1 formed a regulatory network to regulate growth of colorectal cancer. Scale bar = 50 μm. * P
    Figure Legend Snippet: SNHG 1 promotes colorectal cancer progression partly by regulating KLF2 and CDKN2B expression. a Left panel, CCK-8 assays demonstrated that silence of SNHG1 inhibited cancer cell growth. KLF2 knockdown could rescue growth inhibition caused by SNHG1 knockdown in HCT-116 cells. Right panel, CCK-8 assays demonstrated that silence of SNHG1 inhibited cancer cell grow th. CDKN2B (P15) knockdown could rescue growth inhibition caused by SNHG1 knockdown in HCT-116 cells. b EdU assays showed that SNHG1 knockdown inhibited cancer cell proliferation. Co-transfecting KLF2 or CDKN2B siRNAs with SNHG1 siRNAs reversed the decreased proliferation rates in HCT-116 cells. c EdU assays showed that EZH2 knockdown could inhibit proliferation promotion caused by SNHG1 overexpression in HCT-116 cells. d Immunohistochemistry analysis of EZH2, KLF2 and CDKN2B protein levels in colorectal cancer and normal tissues. e Immunohistochemistry analysis of KLF2 and CDKN2B protein levels in tumor tissues formed from SNHG1 knockdown or control cells. f Schematic of the proposed mechanism of SNHG1 in colorectal cancer cells. In the cytoplasm, SNHG1 acts as a ceRNA to sponge miR-154-5p and upregulated the expression of CCND2 (CyclinD2). In the nucleus, SNHG1 is involved in PRC2 mediated epigenetic repression of KLF2 and CDKN2B (P15). KLF2 is also an upstream regulatory factor of CDKN2B. Besides, CDKN2B is a well-studied inhibitor of CCND2. Downstream genes of SNHG1 formed a regulatory network to regulate growth of colorectal cancer. Scale bar = 50 μm. * P

    Techniques Used: Expressing, CCK-8 Assay, Inhibition, Over Expression, Immunohistochemistry

    SNHG1 regulates expression of the miR-154-5p target gene, CCND2. a CCND2 expression was detected by qRT-PCR in SNHG1 siRNAs transfected or SNHG1 siRNAs and miR-154-5p inhibitors co-transfected HCT-116 cells. b CCND2 expression was detected by qRT-PCR in SNHG1 vector transfected or SNHG1 vector and miR-154-5p mimics co-transfected HCT-116 cells. c Western blot analyses of CCND2 expression after knockdown of SNHG1, overexpression of miR-154-5p or knockdown of SNHG1 + inhibition of miR-154-5p in HCT-116 cells. d Dual luciferase reporter assays demonstrated that miR-154-5p overexpression reduced Luc-CCND2 luciferase activity and SNHG1 overexpression abolished miR-154-5p induced reductions in luciferase activity in HCT-116 cells. e CCND2 expression was measured by western blot after silencing of endogenous SNHG1 and transfection with either SNHG1-mut vector, which contains mutations at the putative miR-154-5p binding site, or SNHG1 vector in HCT-116 cells. f The correlation between CCND2 and SNHG1 expression analyzed in 30 paired colorectal cancer samples ( n = 30, r = 0.38, P = 0.036). g EdU assays demonstrated HCT-116 cells proliferation rates after knockdown of SNHG1, knockdown of CCND2 or both knockdown of SNHG1and CCND2. h CCK-8 assays demonstrated that CCND2 knockdown could reverse growth promotion caused by SNHG1 overexpression in HCT-116 cells. i Immunohistochemistry analysis of CCND2 protein levels in tumor tissues formed from SNHG1 knockdown or control cells. j Detection of CCND2 protein levels in colorectal cancer and normal tissues by IHC. Scale bar = 50 μm. * P
    Figure Legend Snippet: SNHG1 regulates expression of the miR-154-5p target gene, CCND2. a CCND2 expression was detected by qRT-PCR in SNHG1 siRNAs transfected or SNHG1 siRNAs and miR-154-5p inhibitors co-transfected HCT-116 cells. b CCND2 expression was detected by qRT-PCR in SNHG1 vector transfected or SNHG1 vector and miR-154-5p mimics co-transfected HCT-116 cells. c Western blot analyses of CCND2 expression after knockdown of SNHG1, overexpression of miR-154-5p or knockdown of SNHG1 + inhibition of miR-154-5p in HCT-116 cells. d Dual luciferase reporter assays demonstrated that miR-154-5p overexpression reduced Luc-CCND2 luciferase activity and SNHG1 overexpression abolished miR-154-5p induced reductions in luciferase activity in HCT-116 cells. e CCND2 expression was measured by western blot after silencing of endogenous SNHG1 and transfection with either SNHG1-mut vector, which contains mutations at the putative miR-154-5p binding site, or SNHG1 vector in HCT-116 cells. f The correlation between CCND2 and SNHG1 expression analyzed in 30 paired colorectal cancer samples ( n = 30, r = 0.38, P = 0.036). g EdU assays demonstrated HCT-116 cells proliferation rates after knockdown of SNHG1, knockdown of CCND2 or both knockdown of SNHG1and CCND2. h CCK-8 assays demonstrated that CCND2 knockdown could reverse growth promotion caused by SNHG1 overexpression in HCT-116 cells. i Immunohistochemistry analysis of CCND2 protein levels in tumor tissues formed from SNHG1 knockdown or control cells. j Detection of CCND2 protein levels in colorectal cancer and normal tissues by IHC. Scale bar = 50 μm. * P

    Techniques Used: Expressing, Quantitative RT-PCR, Transfection, Plasmid Preparation, Western Blot, Over Expression, Inhibition, Luciferase, Activity Assay, Binding Assay, CCK-8 Assay, Immunohistochemistry

    SNHG1 participates in epigenetic repression of KLF2 and CDKN2B by interacting with PRC2. a GSEA showed a significant correlation between the SNHG1 and genes in PRC2 related pathway. b Scatter plot showing the expression relationship among SNHG1, EZH2, SUZ12 and EED in colorectal tumor tissues from TCGA database. The upper right squares show the Pearson correlation between each other. c RIPs experiments for EZH2, SUZ12 and EED were performed and the coprecipitated RNA was subjected to qRT-PCR for SNHG1. GAPDH was employed as a negative control. d RNA pull-down was used to examine the association of SNHG1 and EZH2. AR binding to HuR was used as a positive control. e PRC2 target genes expression was detected by qRT-PCR in SNHG1 siRNAs transfected HCT-116 cells. f CDKN2B and KLF2 expression was detected by qRT-PCR in SNHG1 vector transfected or SNHG1 vector and EZH2 siRNAs co-transfected CRC cells. g CDKN2B and KLF2 protein levels were detected by western blot in indicated conditions. h ChIP assays were performed to detect EZH2 and H3K27me3 occupancy in the CDKN2B promoter region. i ChIP assays were performed to detect EZH2 and H3K27me3 occupancy in the KLF2 promoter region. * P
    Figure Legend Snippet: SNHG1 participates in epigenetic repression of KLF2 and CDKN2B by interacting with PRC2. a GSEA showed a significant correlation between the SNHG1 and genes in PRC2 related pathway. b Scatter plot showing the expression relationship among SNHG1, EZH2, SUZ12 and EED in colorectal tumor tissues from TCGA database. The upper right squares show the Pearson correlation between each other. c RIPs experiments for EZH2, SUZ12 and EED were performed and the coprecipitated RNA was subjected to qRT-PCR for SNHG1. GAPDH was employed as a negative control. d RNA pull-down was used to examine the association of SNHG1 and EZH2. AR binding to HuR was used as a positive control. e PRC2 target genes expression was detected by qRT-PCR in SNHG1 siRNAs transfected HCT-116 cells. f CDKN2B and KLF2 expression was detected by qRT-PCR in SNHG1 vector transfected or SNHG1 vector and EZH2 siRNAs co-transfected CRC cells. g CDKN2B and KLF2 protein levels were detected by western blot in indicated conditions. h ChIP assays were performed to detect EZH2 and H3K27me3 occupancy in the CDKN2B promoter region. i ChIP assays were performed to detect EZH2 and H3K27me3 occupancy in the KLF2 promoter region. * P

    Techniques Used: Expressing, Quantitative RT-PCR, Negative Control, Binding Assay, Positive Control, Transfection, Plasmid Preparation, Western Blot, Chromatin Immunoprecipitation

    SP1 activates SNHG1 transcription in colorectal cancer cells. a Analysis of SP1 ChIP-seq, H3K4me3 ChIP-seq and DnaseI-seq data of HCT-116 cells in the SNHG1 locus. b SNHG1 expression was detected by qRT-PCR in HCT-116 and HCT-8 cells transfected with SP siRNAs or the SP1 vector. c The correlation between SP1 and SNHG1 expression analyzed in 30 paired colorectal cancer samples ( n = 30, r = 0.38, P = 0.03). d ChIP assays were performed to detect SP1 occupancy at the SNHG1 promoter region, α-Satellite and DHFR were employed as negative and positive control respectively for SP1 ChIP assays. e Dual luciferase reporter assays were used to determine the SP1 binding sites on the SNHG1 promoter region. The upper left corner of the picture was SP1 binding motif provided by the JASPAR CORE database. * P
    Figure Legend Snippet: SP1 activates SNHG1 transcription in colorectal cancer cells. a Analysis of SP1 ChIP-seq, H3K4me3 ChIP-seq and DnaseI-seq data of HCT-116 cells in the SNHG1 locus. b SNHG1 expression was detected by qRT-PCR in HCT-116 and HCT-8 cells transfected with SP siRNAs or the SP1 vector. c The correlation between SP1 and SNHG1 expression analyzed in 30 paired colorectal cancer samples ( n = 30, r = 0.38, P = 0.03). d ChIP assays were performed to detect SP1 occupancy at the SNHG1 promoter region, α-Satellite and DHFR were employed as negative and positive control respectively for SP1 ChIP assays. e Dual luciferase reporter assays were used to determine the SP1 binding sites on the SNHG1 promoter region. The upper left corner of the picture was SP1 binding motif provided by the JASPAR CORE database. * P

    Techniques Used: Chromatin Immunoprecipitation, Expressing, Quantitative RT-PCR, Transfection, Plasmid Preparation, Positive Control, Luciferase, Binding Assay

    SNHG1 regulates colorectal cancer cell proliferation and apoptosis. a Results of gene set enrichment analysis (GSEA) were plotted to visualize the correlation between the expression of SNHG1 and cell cycle and DNA repair gene signatures in TCGA cohort. b EdU assays were used to determine the cell proliferation ability of si-SNHG1 transfected cells. c Flow cytometric cell cycle distribution assays to detect the proportion of colorectal cancer cell cells in G1, S, and G2/M phases after transfection with SNHG1 siRNAs. d The cell cycle related proteins CyclinD1, CDK4, CDK6, and CyclinD2 were detected by western blot following SNHG1 silencing. e The effect of SNHG1 knockdown on cell apoptosis was analyzed by flow cytometric cell apoptosis assays. f Apoptosis related proteins Caspase-3, cleaved Caspase-3, PARP, cleaved PARP and Bax were detected by western blot after SNHG1 knockdown. Scale bar = 50 μm. ** P
    Figure Legend Snippet: SNHG1 regulates colorectal cancer cell proliferation and apoptosis. a Results of gene set enrichment analysis (GSEA) were plotted to visualize the correlation between the expression of SNHG1 and cell cycle and DNA repair gene signatures in TCGA cohort. b EdU assays were used to determine the cell proliferation ability of si-SNHG1 transfected cells. c Flow cytometric cell cycle distribution assays to detect the proportion of colorectal cancer cell cells in G1, S, and G2/M phases after transfection with SNHG1 siRNAs. d The cell cycle related proteins CyclinD1, CDK4, CDK6, and CyclinD2 were detected by western blot following SNHG1 silencing. e The effect of SNHG1 knockdown on cell apoptosis was analyzed by flow cytometric cell apoptosis assays. f Apoptosis related proteins Caspase-3, cleaved Caspase-3, PARP, cleaved PARP and Bax were detected by western blot after SNHG1 knockdown. Scale bar = 50 μm. ** P

    Techniques Used: Expressing, Transfection, Flow Cytometry, Western Blot

    SNHG1 affects colorectal cancer cells growth. a SNHG1 expression was detected by qRT-PCR in HCT-116 and HCT-8 cells transfected with two SNHG1 siRNAs. b HCT-116 and HCT-8 cells transfected with SNHG1 siRNAs were subjected to the CCK-8 assay after transfection. c HCT-116 and HCT-8 cells transfected with SNHG1 siRNAs were seeded onto 6-well plates. The number of colonies was counted on the 14th day after seeding. d Representative images of mice bearing tumors from empty vector, sh-SNHG1#1 vector and SNHG1 vector groups, and the tumor volume growth curves after injections in different groups. e SNHG1 expression was detected in tumors from different groups of mice using qRT-PCR. f Representative images of hematoxylin and eosin (HE) staining and Ki67 immunostaining of tumor samples from different groups. Scale bar = 50 μm. * P
    Figure Legend Snippet: SNHG1 affects colorectal cancer cells growth. a SNHG1 expression was detected by qRT-PCR in HCT-116 and HCT-8 cells transfected with two SNHG1 siRNAs. b HCT-116 and HCT-8 cells transfected with SNHG1 siRNAs were subjected to the CCK-8 assay after transfection. c HCT-116 and HCT-8 cells transfected with SNHG1 siRNAs were seeded onto 6-well plates. The number of colonies was counted on the 14th day after seeding. d Representative images of mice bearing tumors from empty vector, sh-SNHG1#1 vector and SNHG1 vector groups, and the tumor volume growth curves after injections in different groups. e SNHG1 expression was detected in tumors from different groups of mice using qRT-PCR. f Representative images of hematoxylin and eosin (HE) staining and Ki67 immunostaining of tumor samples from different groups. Scale bar = 50 μm. * P

    Techniques Used: Expressing, Quantitative RT-PCR, Transfection, CCK-8 Assay, Mouse Assay, Plasmid Preparation, Staining, Immunostaining

    17) Product Images from "Roscovitine synergizes with conventional chemo-therapeutic drugs to induce efficient apoptosis of human colorectal cancer cells"

    Article Title: Roscovitine synergizes with conventional chemo-therapeutic drugs to induce efficient apoptosis of human colorectal cancer cells

    Journal:

    doi: 10.3748/wjg.14.5162

    Modulation of 5-FU cytotoxicity on human colorectal cancer cells by combination with the CDKI Rosco
    Figure Legend Snippet: Modulation of 5-FU cytotoxicity on human colorectal cancer cells by combination with the CDKI Rosco

    Techniques Used:

    Potentiation of 5-FU anticancer effect on human colorectal cancer cell lines by combination with cyclin dependent kinase inhibitor roscovitine. A: Human colorectal cancer cell lines were treated with 5-FU (10 -9 -10 -4 mol/L), and the combination of 5-FU
    Figure Legend Snippet: Potentiation of 5-FU anticancer effect on human colorectal cancer cell lines by combination with cyclin dependent kinase inhibitor roscovitine. A: Human colorectal cancer cell lines were treated with 5-FU (10 -9 -10 -4 mol/L), and the combination of 5-FU

    Techniques Used:

    Modulation of 5-FU cytotoxicity on human colorectal cancer cells by combination with the CDKI Rosco
    Figure Legend Snippet: Modulation of 5-FU cytotoxicity on human colorectal cancer cells by combination with the CDKI Rosco

    Techniques Used:

    Modulation of vinblastine cytotoxicity on human colorectal cancer cells by combination with CDKI Rosco
    Figure Legend Snippet: Modulation of vinblastine cytotoxicity on human colorectal cancer cells by combination with CDKI Rosco

    Techniques Used:

    Modulation of taxol cytotoxicity on human colorectal cancer cells by combination with CDKI Rosco
    Figure Legend Snippet: Modulation of taxol cytotoxicity on human colorectal cancer cells by combination with CDKI Rosco

    Techniques Used:

    Modulation of vinblastine cytotoxicity on human colorectal cancer cells by combination with CDKI Rosco
    Figure Legend Snippet: Modulation of vinblastine cytotoxicity on human colorectal cancer cells by combination with CDKI Rosco

    Techniques Used:

    Potentiation of anticancer effect of taxol on human colorectal cancer cell lines by combination with cyclin dependent kinase inhibitor roscovitine. A: Human colorectal cancer cell lines were treated with taxol (10 -11 -10 -6 mol/L), or the combination of
    Figure Legend Snippet: Potentiation of anticancer effect of taxol on human colorectal cancer cell lines by combination with cyclin dependent kinase inhibitor roscovitine. A: Human colorectal cancer cell lines were treated with taxol (10 -11 -10 -6 mol/L), or the combination of

    Techniques Used:

    Modulation of taxol cytotoxicity on human colorectal cancer cells by combination with CDKI Rosco
    Figure Legend Snippet: Modulation of taxol cytotoxicity on human colorectal cancer cells by combination with CDKI Rosco

    Techniques Used:

    Potentiation of vinblastine anticancer effect on human colorectal cancer cell lines by combination with cyclin dependent kinase inhibitor roscovitine. A: Human colorectal cancer cell lines were treated with vinblastine (10 -12 -10 -7 mol/L) and the combination
    Figure Legend Snippet: Potentiation of vinblastine anticancer effect on human colorectal cancer cell lines by combination with cyclin dependent kinase inhibitor roscovitine. A: Human colorectal cancer cell lines were treated with vinblastine (10 -12 -10 -7 mol/L) and the combination

    Techniques Used:

    Potentiation of doxorubicin anticancer effect on human colorectal cancer cell lines by combination with cyclin dependent kinase inhibitor roscovitine. A: Human colorectal cancer cell lines were treated with doxorubicin (10 -11 -10 -6 mol/L), and the combination
    Figure Legend Snippet: Potentiation of doxorubicin anticancer effect on human colorectal cancer cell lines by combination with cyclin dependent kinase inhibitor roscovitine. A: Human colorectal cancer cell lines were treated with doxorubicin (10 -11 -10 -6 mol/L), and the combination

    Techniques Used:

    Modulation of doxorubicin cytotoxicity on human colorectal cancer cells by combination with CDKI Rosco
    Figure Legend Snippet: Modulation of doxorubicin cytotoxicity on human colorectal cancer cells by combination with CDKI Rosco

    Techniques Used:

    Time and dose dependent effect of roscovitine on the proliferation of human colorectal cancer cell lines. Human colorectal cancer cell lines SW48 (A), SW1116 (B) and SW837 (C) were plated (27 × 10 3 cells/well) into 96-well plates and incubated
    Figure Legend Snippet: Time and dose dependent effect of roscovitine on the proliferation of human colorectal cancer cell lines. Human colorectal cancer cell lines SW48 (A), SW1116 (B) and SW837 (C) were plated (27 × 10 3 cells/well) into 96-well plates and incubated

    Techniques Used: Incubation

    Modulation of doxorubicin cytotoxicity on human colorectal cancer cells by combination with CDKI Rosco
    Figure Legend Snippet: Modulation of doxorubicin cytotoxicity on human colorectal cancer cells by combination with CDKI Rosco

    Techniques Used:

    18) Product Images from "STAT3 Is Necessary for Proliferation and Survival in Colon Cancer–Initiating Cells"

    Article Title: STAT3 Is Necessary for Proliferation and Survival in Colon Cancer–Initiating Cells

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-10-4660

    The ALDH+ /CD133+ subpopulation of colorectal cancer cells express higher levels of STAT3 phosphorylation compared with the ALDH− /CD133− subpopulation
    Figure Legend Snippet: The ALDH+ /CD133+ subpopulation of colorectal cancer cells express higher levels of STAT3 phosphorylation compared with the ALDH− /CD133− subpopulation

    Techniques Used:

    A, LLL12 (5 μmol/L) and Stattic (10 μmol/L) decreased the percentage of ALDH + /CD133 + subpopulation. However, 10 μmol/L doxorubicin or 5-Fu increased the percentage of ALDH + /CD133 + colorectal cancer–initiating cells ( −
    Figure Legend Snippet: A, LLL12 (5 μmol/L) and Stattic (10 μmol/L) decreased the percentage of ALDH + /CD133 + subpopulation. However, 10 μmol/L doxorubicin or 5-Fu increased the percentage of ALDH + /CD133 + colorectal cancer–initiating cells ( −

    Techniques Used:

    19) Product Images from "Pharmacological Inhibition of TFF3 Enhances Sensitivity of CMS4 Colorectal Carcinoma to 5-Fluorouracil through Inhibition of p44/42 MAPK"

    Article Title: Pharmacological Inhibition of TFF3 Enhances Sensitivity of CMS4 Colorectal Carcinoma to 5-Fluorouracil through Inhibition of p44/42 MAPK

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20246215

    TFF3 activates the ERK1/2 (p44/42 MAPK) pathway in CMS4 CRC cells. ( A ) Western blot analysis of phosphorylated and total MAPKs in Caco2-Vec and Caco2-TFF3 cells and SW620-siSC and SW620-siTFF3 cells. β-ACTIN was used as input control. ( B ) Caco2 and SW620 cells were treated with the indicated concentrations of AMPC (with DMSO as vehicle) for 24 h. Levels of p-ERK1/2 and expression of total ERK1/2 were detected by western blot analysis. β-ACTIN was used as an input control. ( C ) Caco2-Vec and Caco2-TFF3 cells were treated with the indicated concentrations of MEK inhibitor (CI1040) or DMSO (vehicle control) for 24 h. Levels of p-ERK1/2 and expression of ERK1/2 were detected by western blot analysis. β-ACTIN was used as an input control. ( D ) Annexin V/PI staining analysis was performed to determine apoptosis in Caco2 stable cells. ***, p
    Figure Legend Snippet: TFF3 activates the ERK1/2 (p44/42 MAPK) pathway in CMS4 CRC cells. ( A ) Western blot analysis of phosphorylated and total MAPKs in Caco2-Vec and Caco2-TFF3 cells and SW620-siSC and SW620-siTFF3 cells. β-ACTIN was used as input control. ( B ) Caco2 and SW620 cells were treated with the indicated concentrations of AMPC (with DMSO as vehicle) for 24 h. Levels of p-ERK1/2 and expression of total ERK1/2 were detected by western blot analysis. β-ACTIN was used as an input control. ( C ) Caco2-Vec and Caco2-TFF3 cells were treated with the indicated concentrations of MEK inhibitor (CI1040) or DMSO (vehicle control) for 24 h. Levels of p-ERK1/2 and expression of ERK1/2 were detected by western blot analysis. β-ACTIN was used as an input control. ( D ) Annexin V/PI staining analysis was performed to determine apoptosis in Caco2 stable cells. ***, p

    Techniques Used: Western Blot, Expressing, Staining

    TFF3 promoted cancer stem cell–like behaviour in mesenchymal colorectal carcinoma (CMS4 CRC) cells. ( A ) qPCR analysis of TFF3 mRNA in Caco2 and SW620 cells was normalized to β-ACTIN . TFF3 expression in monolayer vs. spheroid culture is presented as fold change. ( B ) Western blot analysis for protein expression of TFF3 in monolayer or spheroid culture. β-ACTIN was used as input control. ( C ) Spheroid formation assay. Caco2-Vec and Caco2-TFF3 cells were seeded in ultra-low attachment plates in spheroid growth media for 10 days. Results are presented as percentages relative to the respective control cells (numbers of colonizes are shown). Representative images are shown. Scale bar: 200 μm. ( D ) Spheroid formation of SW620-siSC and SW620-siTFF3 cells as in ( C ). ( E ) The ALDH1+ cell population was determined in Caco2 cells using the ALDEFLUOR assay. Cells were incubated with Aldefluor substrate (BAAA, BoDIPY ® -aminoacetaldehyde) to define ALDH1 positivity and diethylaminobenzaldehyde (DEAB), a specific inhibitor of ALDH1, was used as control to establish the baseline fluorescence. The percentage of the ALDH1+ cells are plotted. ( F ) The ALDH1+ cell population was determined in SW620-siSC and SW620-siTFF3 cells using ALDEFLUOR assay as in ( E ). Data are expressed as mean ±SD. *, p
    Figure Legend Snippet: TFF3 promoted cancer stem cell–like behaviour in mesenchymal colorectal carcinoma (CMS4 CRC) cells. ( A ) qPCR analysis of TFF3 mRNA in Caco2 and SW620 cells was normalized to β-ACTIN . TFF3 expression in monolayer vs. spheroid culture is presented as fold change. ( B ) Western blot analysis for protein expression of TFF3 in monolayer or spheroid culture. β-ACTIN was used as input control. ( C ) Spheroid formation assay. Caco2-Vec and Caco2-TFF3 cells were seeded in ultra-low attachment plates in spheroid growth media for 10 days. Results are presented as percentages relative to the respective control cells (numbers of colonizes are shown). Representative images are shown. Scale bar: 200 μm. ( D ) Spheroid formation of SW620-siSC and SW620-siTFF3 cells as in ( C ). ( E ) The ALDH1+ cell population was determined in Caco2 cells using the ALDEFLUOR assay. Cells were incubated with Aldefluor substrate (BAAA, BoDIPY ® -aminoacetaldehyde) to define ALDH1 positivity and diethylaminobenzaldehyde (DEAB), a specific inhibitor of ALDH1, was used as control to establish the baseline fluorescence. The percentage of the ALDH1+ cells are plotted. ( F ) The ALDH1+ cell population was determined in SW620-siSC and SW620-siTFF3 cells using ALDEFLUOR assay as in ( E ). Data are expressed as mean ±SD. *, p

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Tube Formation Assay, Incubation, Fluorescence

    20) Product Images from "Epigenetic inactivation of the NORE1 gene correlates with malignant progression of colorectal tumors"

    Article Title: Epigenetic inactivation of the NORE1 gene correlates with malignant progression of colorectal tumors

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-10-577

    Methylation status of CpG sites in the NORE1A promoter . A map of the CpG sites within the proximal region of the NORE1A promoter (A). CpGs sites are represented by vertical lines. The transcription start site is indicated by an arrow at +1. Nucleotide sequences of NORE1A promoter region examined by bisulfite DNA sequencing analysis (B). Thirty one CpG sites (bold) examined are numbered 1-31 and primers for PCR were indicated. The ATG start codon is indicated (boxed). Methylation status of 31 CpG sites in the NORE1A promoter and its association with mRNA level in colorectal cancer cell lines (C). The promoter region comprised of 31 CpGs was amplified by PCR. The PCR products were cloned and 5 plasmid clones were sequenced for each cell line. Percent methylation was determined by the number of alleles containing a methylated CpG at each position. Black, gray, and white squares represent complete methylation (4-5 clones), partial methylation (1-3 clones), and unmethylation (0 clone), respectively. Comparison of CpG sites methylation between primary tumors and adjacent noncancerous tissues (D). N, noncancerous tissue; T, tumor tissue; P, patient.
    Figure Legend Snippet: Methylation status of CpG sites in the NORE1A promoter . A map of the CpG sites within the proximal region of the NORE1A promoter (A). CpGs sites are represented by vertical lines. The transcription start site is indicated by an arrow at +1. Nucleotide sequences of NORE1A promoter region examined by bisulfite DNA sequencing analysis (B). Thirty one CpG sites (bold) examined are numbered 1-31 and primers for PCR were indicated. The ATG start codon is indicated (boxed). Methylation status of 31 CpG sites in the NORE1A promoter and its association with mRNA level in colorectal cancer cell lines (C). The promoter region comprised of 31 CpGs was amplified by PCR. The PCR products were cloned and 5 plasmid clones were sequenced for each cell line. Percent methylation was determined by the number of alleles containing a methylated CpG at each position. Black, gray, and white squares represent complete methylation (4-5 clones), partial methylation (1-3 clones), and unmethylation (0 clone), respectively. Comparison of CpG sites methylation between primary tumors and adjacent noncancerous tissues (D). N, noncancerous tissue; T, tumor tissue; P, patient.

    Techniques Used: Methylation, DNA Sequencing, Polymerase Chain Reaction, Amplification, Clone Assay, Plasmid Preparation

    Quantitative analysis of NORE1 mRNA levels in colorectal cancer cell lines and tissue specimens . Expression levels of NORE1A and NORE1B in colorectal tissues and cell lines (A). Quantitation was achieved by densitometric scanning of RT-PCR products in ethidium bromide-stained gels. Absolute area integrations of the curves representing each specimen were compared after adjustment for GAPDH . Data represent means of triplicate assays (Bars, SD) Bar indicates the mean expression level of each specimen group. Expression status of NORE1A and NORE1B in matched tumor sets (B). Expression levels of NORE1A and NORE1B were compared between cancerous and adjacent noncancerous tissues from the same patients. Semi-quantitative RT-PCR was repeated at least three times for each specimen (*, P
    Figure Legend Snippet: Quantitative analysis of NORE1 mRNA levels in colorectal cancer cell lines and tissue specimens . Expression levels of NORE1A and NORE1B in colorectal tissues and cell lines (A). Quantitation was achieved by densitometric scanning of RT-PCR products in ethidium bromide-stained gels. Absolute area integrations of the curves representing each specimen were compared after adjustment for GAPDH . Data represent means of triplicate assays (Bars, SD) Bar indicates the mean expression level of each specimen group. Expression status of NORE1A and NORE1B in matched tumor sets (B). Expression levels of NORE1A and NORE1B were compared between cancerous and adjacent noncancerous tissues from the same patients. Semi-quantitative RT-PCR was repeated at least three times for each specimen (*, P

    Techniques Used: Expressing, Quantitation Assay, Reverse Transcription Polymerase Chain Reaction, Staining, Quantitative RT-PCR

    Association of NORE1 alteration with tumor progression . Correlation of NORE1 reduction with tumor stage and grade (A). Expression levels of NORE1A and NORE1B were compared between stages I, II, and III tumors, and between well differentiated (WD), moderately differentiated (MD), and poorly differentiated (PD) tumors. No association of NORE1A and NORE1B expression with K-Ras mutation in primary colorectal tumors (B). The mutational status of K-Ras in 80 primary carcinoma tissues was determined by SSCP and DNA sequencing analysis.
    Figure Legend Snippet: Association of NORE1 alteration with tumor progression . Correlation of NORE1 reduction with tumor stage and grade (A). Expression levels of NORE1A and NORE1B were compared between stages I, II, and III tumors, and between well differentiated (WD), moderately differentiated (MD), and poorly differentiated (PD) tumors. No association of NORE1A and NORE1B expression with K-Ras mutation in primary colorectal tumors (B). The mutational status of K-Ras in 80 primary carcinoma tissues was determined by SSCP and DNA sequencing analysis.

    Techniques Used: Expressing, Mutagenesis, DNA Sequencing

    Genomic status of NORE1 and re-activation by 5-Aza-dC . Genomic level of NORE1 in colorectal cancer cell lines (A). The NORE1 gene was amplified by genomic PCR using intron-specific primer sets. Ten μl of the PCR products were resolved on a 2% agarose gel. GAPDH was used as an endogenous control. Comparison of NORE1 gene level between tumor and adjacent noncancerous tissues (B). Effect of 5-Aza-dC and TSA on NORE1A and NORE1B mRNA expression (C). Caco-2 and Colo320 cells were treated with increasing doses of 5-Aza-dC for 4 days. For combined treatment with TSA, the cells were incubated with 5-Aza-dC for 48 h and then exposed to TSA (250 ng/ml) for 48 h. Elevation of NORE1A and NORE1B mRNA expression in low expressor cell lines following 5-Aza-dC treatment (D). Cancer cell lines with low NORE1 level were treated with 5-Aza-dC (5 μM) for 5 days and NORE1A and NORE1B expression was evaluated by RT-PCR analysis. C, untreated control; T, 5-Aza-dC treated.
    Figure Legend Snippet: Genomic status of NORE1 and re-activation by 5-Aza-dC . Genomic level of NORE1 in colorectal cancer cell lines (A). The NORE1 gene was amplified by genomic PCR using intron-specific primer sets. Ten μl of the PCR products were resolved on a 2% agarose gel. GAPDH was used as an endogenous control. Comparison of NORE1 gene level between tumor and adjacent noncancerous tissues (B). Effect of 5-Aza-dC and TSA on NORE1A and NORE1B mRNA expression (C). Caco-2 and Colo320 cells were treated with increasing doses of 5-Aza-dC for 4 days. For combined treatment with TSA, the cells were incubated with 5-Aza-dC for 48 h and then exposed to TSA (250 ng/ml) for 48 h. Elevation of NORE1A and NORE1B mRNA expression in low expressor cell lines following 5-Aza-dC treatment (D). Cancer cell lines with low NORE1 level were treated with 5-Aza-dC (5 μM) for 5 days and NORE1A and NORE1B expression was evaluated by RT-PCR analysis. C, untreated control; T, 5-Aza-dC treated.

    Techniques Used: Activation Assay, Amplification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Expressing, Incubation, Reverse Transcription Polymerase Chain Reaction

    Expression status of NORE1A and NORE1B in human colorectal tissues and cancer cell lines . Expression of NORE1A and NORE1B mRNA in normal colorectal epithelial tissues (A). Semi-quantitative RT-PCR was performed using exon-specific primers and 10 μl of the PCR products were resolved on a 2% agarose gel. GAPDH was used as an endogenous control. Frequent reduction of NORE1A and NORE1B expression in colorectal cancer cell lines (B). An immunoblting assay was performed using NORE1A-specific antibody and chemiluminescence detection. Expression of NORE1A and NORE1B in primary colorectal carcinoma tissues (C). Tumor-specific reduction of NORE1A and NORE1B expression (D). NORE1 expression in tumor (T) and adjacent noncancerous (N) tissues were compared using matched tissue sets obtained from the same patients (P).
    Figure Legend Snippet: Expression status of NORE1A and NORE1B in human colorectal tissues and cancer cell lines . Expression of NORE1A and NORE1B mRNA in normal colorectal epithelial tissues (A). Semi-quantitative RT-PCR was performed using exon-specific primers and 10 μl of the PCR products were resolved on a 2% agarose gel. GAPDH was used as an endogenous control. Frequent reduction of NORE1A and NORE1B expression in colorectal cancer cell lines (B). An immunoblting assay was performed using NORE1A-specific antibody and chemiluminescence detection. Expression of NORE1A and NORE1B in primary colorectal carcinoma tissues (C). Tumor-specific reduction of NORE1A and NORE1B expression (D). NORE1 expression in tumor (T) and adjacent noncancerous (N) tissues were compared using matched tissue sets obtained from the same patients (P).

    Techniques Used: Expressing, Quantitative RT-PCR, Polymerase Chain Reaction, Agarose Gel Electrophoresis

    21) Product Images from "Tumor-Derived Exosome-Educated Hepatic Stellate Cells Regulate Lactate Metabolism of Hypoxic Colorectal Tumor Cells via the IL-6/STAT3 Pathway to Confer Drug Resistance"

    Article Title: Tumor-Derived Exosome-Educated Hepatic Stellate Cells Regulate Lactate Metabolism of Hypoxic Colorectal Tumor Cells via the IL-6/STAT3 Pathway to Confer Drug Resistance

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S253485

    P-ERK and p-AKT in tumor-derived exosomes regulate IL-6 secretion. ( A ) ELISA was used to detect the concentration of IL-6 in the cultured medium of HSCs and aHSCs (LoVo and HCT116 exosome-treated HSCs). ( B ) PCR and Western blotting for the IL-6 expression of HSCs and tumor-derived exosomes pre-treated HSCs. ( C ) IHC was performed to analyze α-SMA and IL-6 in the liver metastases of colorectal cancer patients. ( D ) Western blotting analysis of the expression level of p65 in the cytoplasm and nucleus of HSCs and aHSCs. ( E ) ELISA was used to detect the concentration of IL-6 in the medium of HSCs and aHSCs, with or without NF-κB inhibitors (BAY11-7082). ( F ) Western blotting analysis of the expression of p-ERK, ERK, p-AKT and AKT in HSCs and aHSCs. ( G ) Western blotting analysis of the expression of p-ERK, ERK, p-AKT and AKT in the exosomes of LoVo and HCT116. ( H ) ELISA was used to detect the concentration of IL-6 in the cultured medium of HSCs and aHSCs treated with MK226 and U0126. N=3; ns, no significant difference; *P
    Figure Legend Snippet: P-ERK and p-AKT in tumor-derived exosomes regulate IL-6 secretion. ( A ) ELISA was used to detect the concentration of IL-6 in the cultured medium of HSCs and aHSCs (LoVo and HCT116 exosome-treated HSCs). ( B ) PCR and Western blotting for the IL-6 expression of HSCs and tumor-derived exosomes pre-treated HSCs. ( C ) IHC was performed to analyze α-SMA and IL-6 in the liver metastases of colorectal cancer patients. ( D ) Western blotting analysis of the expression level of p65 in the cytoplasm and nucleus of HSCs and aHSCs. ( E ) ELISA was used to detect the concentration of IL-6 in the medium of HSCs and aHSCs, with or without NF-κB inhibitors (BAY11-7082). ( F ) Western blotting analysis of the expression of p-ERK, ERK, p-AKT and AKT in HSCs and aHSCs. ( G ) Western blotting analysis of the expression of p-ERK, ERK, p-AKT and AKT in the exosomes of LoVo and HCT116. ( H ) ELISA was used to detect the concentration of IL-6 in the cultured medium of HSCs and aHSCs treated with MK226 and U0126. N=3; ns, no significant difference; *P

    Techniques Used: Derivative Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Cell Culture, Polymerase Chain Reaction, Western Blot, Expressing, Immunohistochemistry

    Tumor-derived exosomes induce the activation of HSCs. ( A ) Typical electron microscopy images of exosomes secreted by colorectal cancer (LoVo and HCT116; Scale bar, 100 nm). ( B ) NanoSight particle-tracking analysis of exosomes from LoVo and HCT116. ( C ) The total cell lysates and exosomes of LoVo and HCT116 were analyzed by Western blot using the antibody against exosomal proteins (CD9, TSG101). ( D ) The imaging reveals the delivery of Dio-labeled exosomes (green) to Dil-labeled HSCs (red). The yellow arrows indicate the delivered exosomes, and the representative images are presented (Scale bar, 10 μm). ( E ) Immunofluorescence staining for the α-SMA expression of HSCs and tumor-derived exosomes pretreated HSCs. Scale bar, 20 μm. ( F ) PCR and Western blot analyses of the expression of α-SMA in aHSC. ( G ) IHC was performed to analyze α-SMA in the liver metastases of colorectal cancer patients. N=3; *P
    Figure Legend Snippet: Tumor-derived exosomes induce the activation of HSCs. ( A ) Typical electron microscopy images of exosomes secreted by colorectal cancer (LoVo and HCT116; Scale bar, 100 nm). ( B ) NanoSight particle-tracking analysis of exosomes from LoVo and HCT116. ( C ) The total cell lysates and exosomes of LoVo and HCT116 were analyzed by Western blot using the antibody against exosomal proteins (CD9, TSG101). ( D ) The imaging reveals the delivery of Dio-labeled exosomes (green) to Dil-labeled HSCs (red). The yellow arrows indicate the delivered exosomes, and the representative images are presented (Scale bar, 10 μm). ( E ) Immunofluorescence staining for the α-SMA expression of HSCs and tumor-derived exosomes pretreated HSCs. Scale bar, 20 μm. ( F ) PCR and Western blot analyses of the expression of α-SMA in aHSC. ( G ) IHC was performed to analyze α-SMA in the liver metastases of colorectal cancer patients. N=3; *P

    Techniques Used: Derivative Assay, Activation Assay, Electron Microscopy, Western Blot, Imaging, Labeling, Immunofluorescence, Staining, Expressing, Polymerase Chain Reaction, Immunohistochemistry

    22) Product Images from "LncRNA DLEU1 contributes to colorectal cancer progression via activation of KPNA3"

    Article Title: LncRNA DLEU1 contributes to colorectal cancer progression via activation of KPNA3

    Journal: Molecular Cancer

    doi: 10.1186/s12943-018-0873-2

    DLEU1 interacts with SMARCA1 in CRC cells. a The expression of DLEU1 in cytoplasm and nucleus of HCT8 cells was measured by qRT-PCR. U6 serves as a nuclear control. GAPDH serves as a cytoplasmic control. b SMARCA1 was a potential interactive candidate of DLEU1. Biotin-labeled DLEU1 and intron control were incubated with HCT8 cell lysates, and the enriched products were eluted and separated by SDS-PAGE electrophoresis and silver staining. The differential band appearing in DLEU1 lane was analyzed by mass spectrum. c DLEU1 associated with SMARCA1 as shown by RNA pulldown and Western blot. Biotin-labeled DLEU1 and intron control were added into HCT8 cell lysates, and pulldown assays were performed. d DLEU1 was enriched by SMARCA1 in HCT8 and SW480 cell lysates. e SMARCA1 enriched DLEU1 in HCT8 cell lysates. SMARCA1 antibody was added into cell lysates and enriched RNAs were isolated. Then enriched DLEU1 was analyzed by PCR. f DLEU1 co-localized with SMARCA1 in HCT8 cells as shown by RNA FISH. Green, DLEU1; Red, SMARCA1; Blue, DAPI. Scale bar, 10 μm. g the region of nt 1~ 400 in DLEU1 was important for the interaction with SMARCA1. h DLEU1 (nt 1~ 400) associated with SMARCA1 directly as shown by RNA EMSA assays. i The region of nt 700~ 1050 is indispensable for the function of DLEU1 in colorectal cancer. Overexpression of DLEU1 with deletion of nt 1~ 400 cannot promoted proliferation and metastasis in CC. *** P
    Figure Legend Snippet: DLEU1 interacts with SMARCA1 in CRC cells. a The expression of DLEU1 in cytoplasm and nucleus of HCT8 cells was measured by qRT-PCR. U6 serves as a nuclear control. GAPDH serves as a cytoplasmic control. b SMARCA1 was a potential interactive candidate of DLEU1. Biotin-labeled DLEU1 and intron control were incubated with HCT8 cell lysates, and the enriched products were eluted and separated by SDS-PAGE electrophoresis and silver staining. The differential band appearing in DLEU1 lane was analyzed by mass spectrum. c DLEU1 associated with SMARCA1 as shown by RNA pulldown and Western blot. Biotin-labeled DLEU1 and intron control were added into HCT8 cell lysates, and pulldown assays were performed. d DLEU1 was enriched by SMARCA1 in HCT8 and SW480 cell lysates. e SMARCA1 enriched DLEU1 in HCT8 cell lysates. SMARCA1 antibody was added into cell lysates and enriched RNAs were isolated. Then enriched DLEU1 was analyzed by PCR. f DLEU1 co-localized with SMARCA1 in HCT8 cells as shown by RNA FISH. Green, DLEU1; Red, SMARCA1; Blue, DAPI. Scale bar, 10 μm. g the region of nt 1~ 400 in DLEU1 was important for the interaction with SMARCA1. h DLEU1 (nt 1~ 400) associated with SMARCA1 directly as shown by RNA EMSA assays. i The region of nt 700~ 1050 is indispensable for the function of DLEU1 in colorectal cancer. Overexpression of DLEU1 with deletion of nt 1~ 400 cannot promoted proliferation and metastasis in CC. *** P

    Techniques Used: Expressing, Quantitative RT-PCR, Labeling, Incubation, SDS Page, Electrophoresis, Silver Staining, Western Blot, Isolation, Polymerase Chain Reaction, Fluorescence In Situ Hybridization, Over Expression

    23) Product Images from "SNAIL Induces EMT and Lung Metastasis of Tumours Secreting CXCL2 to Promote the Invasion of M2-Type Immunosuppressed Macrophages in Colorectal Cancer"

    Article Title: SNAIL Induces EMT and Lung Metastasis of Tumours Secreting CXCL2 to Promote the Invasion of M2-Type Immunosuppressed Macrophages in Colorectal Cancer

    Journal: International Journal of Biological Sciences

    doi: 10.7150/ijbs.66854

    CXCL2 activates M2 macrophages and induces the migration and proliferation of tumour cells, leading to SNAIL-mediated tumour proliferation. (A) Effects of SNAIL overexpression or knockout, CXCL2 overexpression or knockout co-culture with TAMs on the ability of proliferation, migration and morphology in CRC cells. (B) Number of DLD1 colonies in A, **, P
    Figure Legend Snippet: CXCL2 activates M2 macrophages and induces the migration and proliferation of tumour cells, leading to SNAIL-mediated tumour proliferation. (A) Effects of SNAIL overexpression or knockout, CXCL2 overexpression or knockout co-culture with TAMs on the ability of proliferation, migration and morphology in CRC cells. (B) Number of DLD1 colonies in A, **, P

    Techniques Used: Migration, Over Expression, Knock-Out, Co-Culture Assay

    SNAIL expression is closely related to macrophage promotion of tumour cell migration and metastasis. (A) Immunohistochemical staining of SNAIL in primary tumours with or without CRC lung metastases, and lung metastases. The scale is 200 µm. (B) The proportion of positive cells per 100 cells, statistical number of A, ***, P
    Figure Legend Snippet: SNAIL expression is closely related to macrophage promotion of tumour cell migration and metastasis. (A) Immunohistochemical staining of SNAIL in primary tumours with or without CRC lung metastases, and lung metastases. The scale is 200 µm. (B) The proportion of positive cells per 100 cells, statistical number of A, ***, P

    Techniques Used: Expressing, Migration, Immunohistochemistry, Staining

    SNAIL-dominated EMT and CXCL2-dominated macrophage infiltration are important features of lung metastases. (A) Colorectal carcinoma in situ and lung metastases heatmap of the 30 DEGs with |log2fold change | > 2, P
    Figure Legend Snippet: SNAIL-dominated EMT and CXCL2-dominated macrophage infiltration are important features of lung metastases. (A) Colorectal carcinoma in situ and lung metastases heatmap of the 30 DEGs with |log2fold change | > 2, P

    Techniques Used: In Situ

    24) Product Images from "Comprehensively Analyzed Macrophage-Regulated Genes Indicate That PSMA2 Promotes Colorectal Cancer Progression"

    Article Title: Comprehensively Analyzed Macrophage-Regulated Genes Indicate That PSMA2 Promotes Colorectal Cancer Progression

    Journal: Frontiers in Oncology

    doi: 10.3389/fonc.2020.618902

    PSMA2 promotes CRC cell proliferation. (A) PSMA2 mRNA is up-regulated in CRC samples using clinical samples. (B) Transfecting with si-PSMA2 significantly reduced the PSMA2 expression in FHC and two colorectal cancer cell lines. (C) PSMA2 silencing declined RKO and HCT-116 cell proliferation using CCK-8 assay. (D) PSMA2 silencing suppressed the colony formation abilities of RKO and HCT-116 cell. * P
    Figure Legend Snippet: PSMA2 promotes CRC cell proliferation. (A) PSMA2 mRNA is up-regulated in CRC samples using clinical samples. (B) Transfecting with si-PSMA2 significantly reduced the PSMA2 expression in FHC and two colorectal cancer cell lines. (C) PSMA2 silencing declined RKO and HCT-116 cell proliferation using CCK-8 assay. (D) PSMA2 silencing suppressed the colony formation abilities of RKO and HCT-116 cell. * P

    Techniques Used: Expressing, CCK-8 Assay

    25) Product Images from "MiR-483 Promotes Colorectal Cancer Cell Biological Progression by Directly Targeting NDRG2 through Regulation of the PI3K/AKT Signaling Pathway and Epithelial-to-Mesenchymal Transition"

    Article Title: MiR-483 Promotes Colorectal Cancer Cell Biological Progression by Directly Targeting NDRG2 through Regulation of the PI3K/AKT Signaling Pathway and Epithelial-to-Mesenchymal Transition

    Journal: Journal of Healthcare Engineering

    doi: 10.1155/2022/4574027

    MiR-483 promoted the EMT and activated the phosphorylation of the PI3K/AKT signaling pathway. (a) The mRNA level of NDRG2 in nonnecrotic colon tissues was higher than that in colorectal cancer tissues. (b) NDRG2 was lowly expressed in colorectal cancer cells than normal cells. (c) Western blot illuminated that miR-483 promoted the EMT through NDRG2. (d) MiR-483 activated the PI3K/AKT pathway. ∗ P
    Figure Legend Snippet: MiR-483 promoted the EMT and activated the phosphorylation of the PI3K/AKT signaling pathway. (a) The mRNA level of NDRG2 in nonnecrotic colon tissues was higher than that in colorectal cancer tissues. (b) NDRG2 was lowly expressed in colorectal cancer cells than normal cells. (c) Western blot illuminated that miR-483 promoted the EMT through NDRG2. (d) MiR-483 activated the PI3K/AKT pathway. ∗ P

    Techniques Used: Western Blot

    MiR-483 enhanced the growth of xenograft in vivo. (a) MiR-483 promoted colorectal cancer growth in vivo. (b) The tumor volume of cells overexpressing miR-483 was bigger than the control group. ∗ P
    Figure Legend Snippet: MiR-483 enhanced the growth of xenograft in vivo. (a) MiR-483 promoted colorectal cancer growth in vivo. (b) The tumor volume of cells overexpressing miR-483 was bigger than the control group. ∗ P

    Techniques Used: In Vivo

    Upregulation of miR-483 predicted poor prognosis of colorectal cancer. (a) The expression of miR-483 was higher in colorectal cancer tissues than in the nonnecrotic colon tissues. (b) Overexpression of miR-483 predicted poor 5-year survival of colorectal cancer patients. ∗ P
    Figure Legend Snippet: Upregulation of miR-483 predicted poor prognosis of colorectal cancer. (a) The expression of miR-483 was higher in colorectal cancer tissues than in the nonnecrotic colon tissues. (b) Overexpression of miR-483 predicted poor 5-year survival of colorectal cancer patients. ∗ P

    Techniques Used: Expressing, Over Expression

    MiR-483 promoted the proliferation and invasion in colorectal cancer cells. (a) MiR-483 was overexpressed in CCD-18Co cells versus LOVO and SW480 cells. (b) The miR-483 mimic or the miR-483 inhibitor was transfected to up- or downregulate miR-483 in LOVO cells. (c) MTT assay revealed that the miR-483 mimic promoted the proliferation in LOVO cells. (d) The invasive ability was enhanced by the miR-483 mimic cell, while it was inhibited by the miR-483 inhibitor in LOVO cells. ∗ P
    Figure Legend Snippet: MiR-483 promoted the proliferation and invasion in colorectal cancer cells. (a) MiR-483 was overexpressed in CCD-18Co cells versus LOVO and SW480 cells. (b) The miR-483 mimic or the miR-483 inhibitor was transfected to up- or downregulate miR-483 in LOVO cells. (c) MTT assay revealed that the miR-483 mimic promoted the proliferation in LOVO cells. (d) The invasive ability was enhanced by the miR-483 mimic cell, while it was inhibited by the miR-483 inhibitor in LOVO cells. ∗ P

    Techniques Used: Transfection, MTT Assay

    26) Product Images from "Berberine and Oligomeric Proanthocyanidins Exhibit Synergistic Efficacy Through Regulation of PI3K-Akt Signaling Pathway in Colorectal Cancer"

    Article Title: Berberine and Oligomeric Proanthocyanidins Exhibit Synergistic Efficacy Through Regulation of PI3K-Akt Signaling Pathway in Colorectal Cancer

    Journal: Frontiers in Oncology

    doi: 10.3389/fonc.2022.855860

    OPCs increased the cellular uptake of BBR in colorectal cancer cell lines. (A) Cellular fluorescence intensity value of BBR following treatment with BBR and combination of BBR and OPCs for 24 hours in RKO and HT29 cells. The average (column) ± SD is indicated (*P
    Figure Legend Snippet: OPCs increased the cellular uptake of BBR in colorectal cancer cell lines. (A) Cellular fluorescence intensity value of BBR following treatment with BBR and combination of BBR and OPCs for 24 hours in RKO and HT29 cells. The average (column) ± SD is indicated (*P

    Techniques Used: Fluorescence

    BBR and OPCs promoted the cell apoptosis in colorectal cancer cell lines. (A) Representative images of cells undergoing apoptosis that stained for annexin V assay in RKO and HT29 cells. The average (column) ± SD is indicated (* P
    Figure Legend Snippet: BBR and OPCs promoted the cell apoptosis in colorectal cancer cell lines. (A) Representative images of cells undergoing apoptosis that stained for annexin V assay in RKO and HT29 cells. The average (column) ± SD is indicated (* P

    Techniques Used: Staining, Annexin V Assay

    The siRNA-based knock-down of MYB inhibited a tumorigenic effect through induction of apoptosis in colorectal cancer cell lines. (A) qRT-PCR analysis of MYB expression in RKO and HT29 cells. MYB expression was knock-downed by siMYB. Relative expression was calculated using β-Actin mRNA expression as an internal control. The average (column) ± SD is indicated (* P
    Figure Legend Snippet: The siRNA-based knock-down of MYB inhibited a tumorigenic effect through induction of apoptosis in colorectal cancer cell lines. (A) qRT-PCR analysis of MYB expression in RKO and HT29 cells. MYB expression was knock-downed by siMYB. Relative expression was calculated using β-Actin mRNA expression as an internal control. The average (column) ± SD is indicated (* P

    Techniques Used: Quantitative RT-PCR, Expressing

    BBR and OPCs modulated multiple cancer-associated pathways. (A, B) Venn-diagram of dysregulated expression ( > 1.5-fold) of the genes following treatment with BBR and OPCs in RKO (A) and HT29 (B) cells. (C, D) The top 10 pathways affected by both BBR and OPCs in RKO (C) and HT29 (D) cells. (E) Heatmap of differentially expressed genes regulated by both BBR and OPCs in PI3K-Akt signaling pathway. (F) qRT-PCR analysis of MYB expression in RKO and HT29 cells following treatment with BBR, OPCs, and their combination for 48 hours. Relative expression was calculated using β-Actin mRNA expression as an internal control. The average (column) ± SD is indicated (*P
    Figure Legend Snippet: BBR and OPCs modulated multiple cancer-associated pathways. (A, B) Venn-diagram of dysregulated expression ( > 1.5-fold) of the genes following treatment with BBR and OPCs in RKO (A) and HT29 (B) cells. (C, D) The top 10 pathways affected by both BBR and OPCs in RKO (C) and HT29 (D) cells. (E) Heatmap of differentially expressed genes regulated by both BBR and OPCs in PI3K-Akt signaling pathway. (F) qRT-PCR analysis of MYB expression in RKO and HT29 cells following treatment with BBR, OPCs, and their combination for 48 hours. Relative expression was calculated using β-Actin mRNA expression as an internal control. The average (column) ± SD is indicated (*P

    Techniques Used: Expressing, Quantitative RT-PCR

    The combination of BBR and OPCs effectively suppressed growth of tumor organoids derived from human colorectal cancers. (A) Schematic protocol of BBR, OPCs, and their combination treatment on tumor organoids derived from human colorectal cancers. (B) Representative images of tumor organoids following treatment with BBR, OPCs, and their combination. Scale bar = 500μm. The average (column) ± SD is indicated (*P
    Figure Legend Snippet: The combination of BBR and OPCs effectively suppressed growth of tumor organoids derived from human colorectal cancers. (A) Schematic protocol of BBR, OPCs, and their combination treatment on tumor organoids derived from human colorectal cancers. (B) Representative images of tumor organoids following treatment with BBR, OPCs, and their combination. Scale bar = 500μm. The average (column) ± SD is indicated (*P

    Techniques Used: Derivative Assay

    BBR and OPCs exert a synergistic anti-tumorigenic effect in colorectal cancer cell lines. (A) CCK-8 assays comparing cell viability following treatment with BBR, OPCs, and their combination for 48 hours in RKO and HT29 cells. Error bars are the mean ± SD. (B) Colony formation assays to assess clonogenicity of CRC cells following treatment with BBR, OPCs, and their combination. The average (column) ± SD is indicated (*P
    Figure Legend Snippet: BBR and OPCs exert a synergistic anti-tumorigenic effect in colorectal cancer cell lines. (A) CCK-8 assays comparing cell viability following treatment with BBR, OPCs, and their combination for 48 hours in RKO and HT29 cells. Error bars are the mean ± SD. (B) Colony formation assays to assess clonogenicity of CRC cells following treatment with BBR, OPCs, and their combination. The average (column) ± SD is indicated (*P

    Techniques Used: CCK-8 Assay

    A schematic illustration of the synergistic anti-cancer effect of BBR and OPCs. This illustration demonstrates treatment of BBR alone (left) and combined treatment of BBR and OPCs (right) in CRC cells. OPCs increased the cellular uptake of BBR in CRC cells and enhanced BBR’s anti-cancer potential through MYB and PI3K-Akt signaling pathway.
    Figure Legend Snippet: A schematic illustration of the synergistic anti-cancer effect of BBR and OPCs. This illustration demonstrates treatment of BBR alone (left) and combined treatment of BBR and OPCs (right) in CRC cells. OPCs increased the cellular uptake of BBR in CRC cells and enhanced BBR’s anti-cancer potential through MYB and PI3K-Akt signaling pathway.

    Techniques Used:

    27) Product Images from "Gilteritinib induces PUMA‐dependent apoptotic cell death via AKT/GSK‐3β/NF‐κB pathway in colorectal cancer cells. Gilteritinib induces PUMA‐dependent apoptotic cell death via AKT/GSK‐3β/NF‐κB pathway in colorectal cancer cells"

    Article Title: Gilteritinib induces PUMA‐dependent apoptotic cell death via AKT/GSK‐3β/NF‐κB pathway in colorectal cancer cells. Gilteritinib induces PUMA‐dependent apoptotic cell death via AKT/GSK‐3β/NF‐κB pathway in colorectal cancer cells

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.14913

    Gilteritinib inhibits cell growth and induces apoptosis in CRC cells. A, The indicated CRC cell lines were treated with increasing concentrations of gilteritinib for 72 h. Cell viability was determined by MTS assay. B, Crystal violet results of HCT116 cells treated with gilteritinib at indicated concentration for 72 h. C, The indicated cell lines were treated with gilteritinib for 24 h at indicated concentrations. Apoptosis was analysed by Annexin V/PI staining followed by flow cytometry. D, The indicated cell lines were treated gilteritinib at indicated concentration. Caspase 3/7 activity was determined by fluorogenic analysis. E, The indicated cell lines were treated 50 nmol/L gilteritinib with or without 10 μmol/L z‐VAD‐fmk pre‐treatment. Apoptosis was analysed by Annexin V/PI staining followed by flow cytometry. F, HCT116 and SW480 cells were treated with 50 nmol/L gilteritinib at indicated time‐point. Cleaved caspase 3 and 9 were analysed by Western blotting. Results in (C), (D) and (E) were expressed as means ± SD of three independent experiments. ** P
    Figure Legend Snippet: Gilteritinib inhibits cell growth and induces apoptosis in CRC cells. A, The indicated CRC cell lines were treated with increasing concentrations of gilteritinib for 72 h. Cell viability was determined by MTS assay. B, Crystal violet results of HCT116 cells treated with gilteritinib at indicated concentration for 72 h. C, The indicated cell lines were treated with gilteritinib for 24 h at indicated concentrations. Apoptosis was analysed by Annexin V/PI staining followed by flow cytometry. D, The indicated cell lines were treated gilteritinib at indicated concentration. Caspase 3/7 activity was determined by fluorogenic analysis. E, The indicated cell lines were treated 50 nmol/L gilteritinib with or without 10 μmol/L z‐VAD‐fmk pre‐treatment. Apoptosis was analysed by Annexin V/PI staining followed by flow cytometry. F, HCT116 and SW480 cells were treated with 50 nmol/L gilteritinib at indicated time‐point. Cleaved caspase 3 and 9 were analysed by Western blotting. Results in (C), (D) and (E) were expressed as means ± SD of three independent experiments. ** P

    Techniques Used: MTS Assay, Concentration Assay, Staining, Flow Cytometry, Activity Assay, Western Blot

    Gilteritinib promotes PUMA induction in CRC cells. A, HCT116 cells were treated with 50 nmol/L gilteritinib at indicated time‐point. PUMA expression was analysed by Western blotting. B, HCT116 cells were treated with 50 nmol/L gilteritinib at indicated time‐point. PUMA mRNA induction by gilteritinib was analysed by real‐time reverse transcriptase (RT) PCR. C, HCT116 cells were treated with 50 nmol/L gilteritinib at indicated time‐point. The expression of indicated Bcl‐2 family members was analysed by Western blotting. D, SW480 cells were treated with 50 nmol/L gilteritinib at indicated time‐point. PUMA expression was analysed by Western blotting. E, Indicated CRC cell lines were treated with 50 nmol/L gilteritinib for 24 hours. PUMA expression was analysed by Western blotting. F, HCT116 cells were treated with 50 nmol/L quizartinib or dovitinib at indicated time‐point. PUMA expression was analysed by Western blotting. Results in (B) were expressed as means ± SD of 3 independent experiments. ** P
    Figure Legend Snippet: Gilteritinib promotes PUMA induction in CRC cells. A, HCT116 cells were treated with 50 nmol/L gilteritinib at indicated time‐point. PUMA expression was analysed by Western blotting. B, HCT116 cells were treated with 50 nmol/L gilteritinib at indicated time‐point. PUMA mRNA induction by gilteritinib was analysed by real‐time reverse transcriptase (RT) PCR. C, HCT116 cells were treated with 50 nmol/L gilteritinib at indicated time‐point. The expression of indicated Bcl‐2 family members was analysed by Western blotting. D, SW480 cells were treated with 50 nmol/L gilteritinib at indicated time‐point. PUMA expression was analysed by Western blotting. E, Indicated CRC cell lines were treated with 50 nmol/L gilteritinib for 24 hours. PUMA expression was analysed by Western blotting. F, HCT116 cells were treated with 50 nmol/L quizartinib or dovitinib at indicated time‐point. PUMA expression was analysed by Western blotting. Results in (B) were expressed as means ± SD of 3 independent experiments. ** P

    Techniques Used: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction

    28) Product Images from "SMAD4 mutations do not preclude epithelial–mesenchymal transition in colorectal cancer"

    Article Title: SMAD4 mutations do not preclude epithelial–mesenchymal transition in colorectal cancer

    Journal: Oncogene

    doi: 10.1038/s41388-021-02128-2

    SMAD4 mutations do not preclude an EMT-like transcriptome in human tumors in vivo. a Assignment of the HT29-ctrl and HT29-Snail1-HA transcriptome samples to consensus molecular subtypes (CMS) of colorectal cancer. For each condition, both replicates (repl.) are plotted individually. n.s.: no significant assignment to one of the four CMS. b SMAD4 mutation frequencies in colon adenocarcinoma (COAD) samples from The Cancer Genome Atlas (TCGA) database. Numbers of total samples and SMAD4 mut frequencies for each CMS as well as in all samples in the dataset (All) are listed. n.d.: no significant CMS classification possible. c, d Single-sample gene set enrichment analysis (ssGSEA) of COAD ( c ) and pancreatic adenocarcinoma (PAAD, ( d )) samples from the TCGA database. Scores for ssGSEA were calculated for each sample based on enrichment of the 61 genes that were upregulated in both HT29-Snail1-HA clones after 6 d of Dox treatment and are contained in at least one of the EMT core signatures (see Fig. 4d, e ). The samples are ordered horizontally according to their ssGSEA score. The yellow/brown color bar below indicates the SMAD4 status of each sample. e, f Correlation heatmaps displaying the mutual correlation of expression levels of the 141 genes that are significantly regulated in both HT29-Snail1-HA clones after 6 d of Dox treatment and are contained in at least one of the EMT core signatures (see Fig. 4d, e ). Tumor transcriptomes were obtained from the TCGA database and samples were divided into two groups according to their SMAD4 mutation status. For e COAD and for f PAAD samples were used. Clustering of genes was achieved by unsupervised hierarchical clustering based on the Euclidean distance. Dotted lines highlight the boundaries between the two principal gene clusters obtained in each case. Turquoise/pink color bars below each heatmap indicate whether a gene is up- or downregulated in HT29-Snail1-HA cells. g Expression levels of EMT transcription factor genes in COAD samples from the TCGA database. Samples were grouped by their CMS and further divided based on their SMAD4 mutation status. Sample numbers for each condition are listed in b . Dots in the box plots indicate average values. Whisker lengths are based on the Tukey method. Outliers are not plotted but were considered for calculating significance. Significance was determined by differential gene expression analysis with the limma package in R/Bioconductor; n.s.: not significant (adj. p -value ≥ 0.05). TPM: transcripts per million.
    Figure Legend Snippet: SMAD4 mutations do not preclude an EMT-like transcriptome in human tumors in vivo. a Assignment of the HT29-ctrl and HT29-Snail1-HA transcriptome samples to consensus molecular subtypes (CMS) of colorectal cancer. For each condition, both replicates (repl.) are plotted individually. n.s.: no significant assignment to one of the four CMS. b SMAD4 mutation frequencies in colon adenocarcinoma (COAD) samples from The Cancer Genome Atlas (TCGA) database. Numbers of total samples and SMAD4 mut frequencies for each CMS as well as in all samples in the dataset (All) are listed. n.d.: no significant CMS classification possible. c, d Single-sample gene set enrichment analysis (ssGSEA) of COAD ( c ) and pancreatic adenocarcinoma (PAAD, ( d )) samples from the TCGA database. Scores for ssGSEA were calculated for each sample based on enrichment of the 61 genes that were upregulated in both HT29-Snail1-HA clones after 6 d of Dox treatment and are contained in at least one of the EMT core signatures (see Fig. 4d, e ). The samples are ordered horizontally according to their ssGSEA score. The yellow/brown color bar below indicates the SMAD4 status of each sample. e, f Correlation heatmaps displaying the mutual correlation of expression levels of the 141 genes that are significantly regulated in both HT29-Snail1-HA clones after 6 d of Dox treatment and are contained in at least one of the EMT core signatures (see Fig. 4d, e ). Tumor transcriptomes were obtained from the TCGA database and samples were divided into two groups according to their SMAD4 mutation status. For e COAD and for f PAAD samples were used. Clustering of genes was achieved by unsupervised hierarchical clustering based on the Euclidean distance. Dotted lines highlight the boundaries between the two principal gene clusters obtained in each case. Turquoise/pink color bars below each heatmap indicate whether a gene is up- or downregulated in HT29-Snail1-HA cells. g Expression levels of EMT transcription factor genes in COAD samples from the TCGA database. Samples were grouped by their CMS and further divided based on their SMAD4 mutation status. Sample numbers for each condition are listed in b . Dots in the box plots indicate average values. Whisker lengths are based on the Tukey method. Outliers are not plotted but were considered for calculating significance. Significance was determined by differential gene expression analysis with the limma package in R/Bioconductor; n.s.: not significant (adj. p -value ≥ 0.05). TPM: transcripts per million.

    Techniques Used: In Vivo, Mutagenesis, Clone Assay, Expressing, Whisker Assay

    29) Product Images from "ABCB5 identifies a therapy-refractory tumor cell population in colorectal cancer patients"

    Article Title: ABCB5 identifies a therapy-refractory tumor cell population in colorectal cancer patients

    Journal: Cancer Research

    doi: 10.1158/0008-5472.CAN-11-0221

    ABCB5 expression in physiological colon and colorectal cancer A , Representative immunohistochemical analysis of ABCB5 protein expression (AP, red, left two panels) or CD133 expression (third panel) in physiological human colon tissue compared to isotype control staining (right panel). Arrows demarcate cells expressing ABCB5 and CD133 in sections of full-length crypts from the lumenal surface of the colon to the crypt bases bordered by underlying connective tissue. B , Representative ABCB5/CD133 coexpression in human colon crypt cells showing basilar staining for ABCB5 (AP, red) and apical staining for CD133 (HRP, brown) in both a cross and lateral section. C , Representative immunofluorescence analysis of ABCB5 (green) and CD133 (red) co-expression in a clinical human colon cancer specimen. The central lumen in the far-right panel (labeled A) is defined at its periphery by apical membrane reactivity for CD133 (labeled B), with associated underlying foci of ABCB5 reactivity (labeled C). D , Left panels: Immunohistochemical analysis of ABCB5 (AP, red) protein expression in well-differentiated versus poorly-differentiated tumor areas of a representative clinical colon cancer. Right panel: ABCB5 protein expression (% positive cells, mean±SEM) in physiological colon ( n =9) versus clinical colon cancer specimens ( n =29), as determined by quantitative image analysis of ABCB5 immunohistochemistry.
    Figure Legend Snippet: ABCB5 expression in physiological colon and colorectal cancer A , Representative immunohistochemical analysis of ABCB5 protein expression (AP, red, left two panels) or CD133 expression (third panel) in physiological human colon tissue compared to isotype control staining (right panel). Arrows demarcate cells expressing ABCB5 and CD133 in sections of full-length crypts from the lumenal surface of the colon to the crypt bases bordered by underlying connective tissue. B , Representative ABCB5/CD133 coexpression in human colon crypt cells showing basilar staining for ABCB5 (AP, red) and apical staining for CD133 (HRP, brown) in both a cross and lateral section. C , Representative immunofluorescence analysis of ABCB5 (green) and CD133 (red) co-expression in a clinical human colon cancer specimen. The central lumen in the far-right panel (labeled A) is defined at its periphery by apical membrane reactivity for CD133 (labeled B), with associated underlying foci of ABCB5 reactivity (labeled C). D , Left panels: Immunohistochemical analysis of ABCB5 (AP, red) protein expression in well-differentiated versus poorly-differentiated tumor areas of a representative clinical colon cancer. Right panel: ABCB5 protein expression (% positive cells, mean±SEM) in physiological colon ( n =9) versus clinical colon cancer specimens ( n =29), as determined by quantitative image analysis of ABCB5 immunohistochemistry.

    Techniques Used: Expressing, Immunohistochemistry, Staining, Immunofluorescence, Labeling

    Response of ABCB5 expression to 5-FU treatment in human colorectal cancer xenografts A , Correlation between ABCB5 expression and 5-FU resistance in human NCI-60 colon cancer cell lines: (1) COLO205, (2) HCT-116, (3) HCT-15, (4) HT-29, (5) HCC-2998, (6) KM12, and (7) SW620. B , Left panel: RT-PCR expression analysis of full-length ABCB5 mRNA (NM_178559) in human SW480 and HT-29 colon cancer cell lines. The human melanoma cell line G3361 was used as a positive control. Right panels: Flow cytometric determination of ABCB5 protein expression in SW480 and HT-29 cells. Bottom panels depict isotype control-stained cells. C , ABCB5 expression in human colorectal cancer xenografts in response to 5-FU treatment. Panels depict (from left to right) representative ABCB5 staining (HRP, brown; nuclei are counterstained with DAPI, blue) or ABCB5 (AP, blue) / TUNEL (HRP, brown) co-staining of SW480 (top rows) and HT-29 (bottom rows) colon cancer xenografts dissected from vehicle control- versus 5-FU treated animals. D , Quantitative ABCB5 protein expression analysis in colon cancer xenografts (% positive cells, mean±SEM) dissected from vehicle-control ( n =13) versus 5-FU-treated specimens ( n =18).
    Figure Legend Snippet: Response of ABCB5 expression to 5-FU treatment in human colorectal cancer xenografts A , Correlation between ABCB5 expression and 5-FU resistance in human NCI-60 colon cancer cell lines: (1) COLO205, (2) HCT-116, (3) HCT-15, (4) HT-29, (5) HCC-2998, (6) KM12, and (7) SW620. B , Left panel: RT-PCR expression analysis of full-length ABCB5 mRNA (NM_178559) in human SW480 and HT-29 colon cancer cell lines. The human melanoma cell line G3361 was used as a positive control. Right panels: Flow cytometric determination of ABCB5 protein expression in SW480 and HT-29 cells. Bottom panels depict isotype control-stained cells. C , ABCB5 expression in human colorectal cancer xenografts in response to 5-FU treatment. Panels depict (from left to right) representative ABCB5 staining (HRP, brown; nuclei are counterstained with DAPI, blue) or ABCB5 (AP, blue) / TUNEL (HRP, brown) co-staining of SW480 (top rows) and HT-29 (bottom rows) colon cancer xenografts dissected from vehicle control- versus 5-FU treated animals. D , Quantitative ABCB5 protein expression analysis in colon cancer xenografts (% positive cells, mean±SEM) dissected from vehicle-control ( n =13) versus 5-FU-treated specimens ( n =18).

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Positive Control, Flow Cytometry, Staining, TUNEL Assay

    Inhibition of tumorigenic growth and 5-FU resistance reversal of human colorectal cancer cells by ABCB5 knockdown (KD) A , Stable HT-29 (top) or SW480 (bottom) ABCB5-KD cells or vector controls were generated using shRNA gene silencing. Confirmation of ABCB5-KD at mRNA (bottom rows, determined by exon-exon RT-PCR), and protein levels (top rows, determined by Western blotting), using GAPDH and αTubulin as controls, respectively. B , Analysis of in vitro growth kinetics of stable HT29 ABCB5 KD (red line) vs. HT-29 control (blue line) cells (top), or SW480 ABCB5 KD (red line) vs. SW480 control (blue line) cells bottom). C , In vivo tumor growth kinetics of stable HT29 ABCB5 KD (red line) vs. HT-29 control (blue line) xenografts (top), or SW480 ABCB5 KD (red line) vs. SW480 control (blue line) xenografts (bottom). D , 5-FU-dependent cell killing for HT-29 control (blue line) vs. HT29 ABCB5 KD cells (red line) (top) and SW480 control (blue line) vs. SW480 ABCB5 KD (red line) cells (bottom) as determined using the MTT assay. Illustrated are surviving cell fractions as a function of 5-FU concentration (nM) for n =6 replicate samples, respectively (*: P
    Figure Legend Snippet: Inhibition of tumorigenic growth and 5-FU resistance reversal of human colorectal cancer cells by ABCB5 knockdown (KD) A , Stable HT-29 (top) or SW480 (bottom) ABCB5-KD cells or vector controls were generated using shRNA gene silencing. Confirmation of ABCB5-KD at mRNA (bottom rows, determined by exon-exon RT-PCR), and protein levels (top rows, determined by Western blotting), using GAPDH and αTubulin as controls, respectively. B , Analysis of in vitro growth kinetics of stable HT29 ABCB5 KD (red line) vs. HT-29 control (blue line) cells (top), or SW480 ABCB5 KD (red line) vs. SW480 control (blue line) cells bottom). C , In vivo tumor growth kinetics of stable HT29 ABCB5 KD (red line) vs. HT-29 control (blue line) xenografts (top), or SW480 ABCB5 KD (red line) vs. SW480 control (blue line) xenografts (bottom). D , 5-FU-dependent cell killing for HT-29 control (blue line) vs. HT29 ABCB5 KD cells (red line) (top) and SW480 control (blue line) vs. SW480 ABCB5 KD (red line) cells (bottom) as determined using the MTT assay. Illustrated are surviving cell fractions as a function of 5-FU concentration (nM) for n =6 replicate samples, respectively (*: P

    Techniques Used: Inhibition, Plasmid Preparation, Generated, shRNA, Reverse Transcription Polymerase Chain Reaction, Western Blot, In Vitro, In Vivo, MTT Assay, Concentration Assay

    30) Product Images from "Depletion of insulin-like growth factor 1 receptor increases radiosensitivity in colorectal cancer"

    Article Title: Depletion of insulin-like growth factor 1 receptor increases radiosensitivity in colorectal cancer

    Journal: Journal of Gastrointestinal Oncology

    doi: 10.21037/jgo-20-210

    BMS-754807 promotes a response in colorectal cancer cells to irradiation. (A,B) Proliferation in SW480 cells treated and untreated with BMS-754807 was significantly reduced in a dose-dependent matter after irradiation.*P
    Figure Legend Snippet: BMS-754807 promotes a response in colorectal cancer cells to irradiation. (A,B) Proliferation in SW480 cells treated and untreated with BMS-754807 was significantly reduced in a dose-dependent matter after irradiation.*P

    Techniques Used: Irradiation

    31) Product Images from "Rapid Multiplex Strip Test for the Detection of Circulating Tumor DNA Mutations for Liquid Biopsy Applications"

    Article Title: Rapid Multiplex Strip Test for the Detection of Circulating Tumor DNA Mutations for Liquid Biopsy Applications

    Journal: Biosensors

    doi: 10.3390/bios12020097

    Application of the multiplex strip test for the detection of cell-free DNA and circulating tumor DNA in blood samples of four healthy individuals and five CRC patients. CRC: colorectal cancer, TZ: test zone, CZ: control zone.
    Figure Legend Snippet: Application of the multiplex strip test for the detection of cell-free DNA and circulating tumor DNA in blood samples of four healthy individuals and five CRC patients. CRC: colorectal cancer, TZ: test zone, CZ: control zone.

    Techniques Used: Multiplex Assay, Stripping Membranes

    32) Product Images from "Adenoviruses with an ?v? integrin targeting moiety in the fiber shaft or the HI-loop increase tumor specificity without compromising antitumor efficacy in magnetic resonance imaging of colorectal cancer metastases"

    Article Title: Adenoviruses with an ?v? integrin targeting moiety in the fiber shaft or the HI-loop increase tumor specificity without compromising antitumor efficacy in magnetic resonance imaging of colorectal cancer metastases

    Journal: Journal of Translational Medicine

    doi: 10.1186/1479-5876-8-80

    Gene transfer to human colorectal cancer cells . Adenoviral vectors targeted for α v β integrins via Arg-Gly-Asp (RGD) modification in the HI loop (Ad5luc1RGD) or the shaft domain (AdTLRGDK) of the fiber showed enhanced gene transfer to human colorectal cancer cell lines. Cells were infected with 1000 VP/cell and luciferase activity was measured 24 hours later. Data is presented as relative light units (RLU) normalized for gene expression of Ad5 control virus AdTL. Each data point represents the mean of three replicates ± SEM.
    Figure Legend Snippet: Gene transfer to human colorectal cancer cells . Adenoviral vectors targeted for α v β integrins via Arg-Gly-Asp (RGD) modification in the HI loop (Ad5luc1RGD) or the shaft domain (AdTLRGDK) of the fiber showed enhanced gene transfer to human colorectal cancer cell lines. Cells were infected with 1000 VP/cell and luciferase activity was measured 24 hours later. Data is presented as relative light units (RLU) normalized for gene expression of Ad5 control virus AdTL. Each data point represents the mean of three replicates ± SEM.

    Techniques Used: Modification, Infection, Luciferase, Activity Assay, Expressing

    Cell killing potency of RGD modified viruses in vitro . Viruses with RGD modification in the capsid display effective killing of colorectal cancer cell lines. Cells were infected with replication competent (WT-RGD, WT-RGDK, WT) or non-replicating (Ad5luc1) viruses and the cell killing potency was assessed with the MTS assay. Data are presented as relative cell viability normalized to mock (growth medium) infected cells. Each data point represents the mean of six replicates ± SEM.
    Figure Legend Snippet: Cell killing potency of RGD modified viruses in vitro . Viruses with RGD modification in the capsid display effective killing of colorectal cancer cell lines. Cells were infected with replication competent (WT-RGD, WT-RGDK, WT) or non-replicating (Ad5luc1) viruses and the cell killing potency was assessed with the MTS assay. Data are presented as relative cell viability normalized to mock (growth medium) infected cells. Each data point represents the mean of six replicates ± SEM.

    Techniques Used: Modification, In Vitro, Infection, MTS Assay

    Antitumor efficacy of RGD modified viruses in the spleen-to-liver colorectal cancer model . Enhanced therapeutic effect of RGD modified replication competent adenoviruses in spleen-to-liver colorectal cancer model. To imitate clinical metastatic colorectal cancer, hepatic tumors were induced in mice by intrasplenic injection of HCT116 colorectal cancer cells. WT, WT-RGD, or WT-RGDK viruses at dose of 3 × 10e10 VP were injected via tail vein in two consecutive days (days 23 and 24). (A) Hepatic tumor growth was followed with MRI thereafter. Relative tumor volumes normalized to the day before virus treatment (day -1) tumor volumes are presented. Each data point represents mean of 2 to 11 measurements ± SEM. *, p
    Figure Legend Snippet: Antitumor efficacy of RGD modified viruses in the spleen-to-liver colorectal cancer model . Enhanced therapeutic effect of RGD modified replication competent adenoviruses in spleen-to-liver colorectal cancer model. To imitate clinical metastatic colorectal cancer, hepatic tumors were induced in mice by intrasplenic injection of HCT116 colorectal cancer cells. WT, WT-RGD, or WT-RGDK viruses at dose of 3 × 10e10 VP were injected via tail vein in two consecutive days (days 23 and 24). (A) Hepatic tumor growth was followed with MRI thereafter. Relative tumor volumes normalized to the day before virus treatment (day -1) tumor volumes are presented. Each data point represents mean of 2 to 11 measurements ± SEM. *, p

    Techniques Used: Modification, Mouse Assay, Injection, Magnetic Resonance Imaging

    33) Product Images from "LncRNA DLEU1 contributes to colorectal cancer progression via activation of KPNA3"

    Article Title: LncRNA DLEU1 contributes to colorectal cancer progression via activation of KPNA3

    Journal: Molecular Cancer

    doi: 10.1186/s12943-018-0873-2

    DLEU1 interacts with SMARCA1 in CRC cells. a The expression of DLEU1 in cytoplasm and nucleus of HCT8 cells was measured by qRT-PCR. U6 serves as a nuclear control. GAPDH serves as a cytoplasmic control. b SMARCA1 was a potential interactive candidate of DLEU1. Biotin-labeled DLEU1 and intron control were incubated with HCT8 cell lysates, and the enriched products were eluted and separated by SDS-PAGE electrophoresis and silver staining. The differential band appearing in DLEU1 lane was analyzed by mass spectrum. c DLEU1 associated with SMARCA1 as shown by RNA pulldown and Western blot. Biotin-labeled DLEU1 and intron control were added into HCT8 cell lysates, and pulldown assays were performed. d DLEU1 was enriched by SMARCA1 in HCT8 and SW480 cell lysates. e SMARCA1 enriched DLEU1 in HCT8 cell lysates. SMARCA1 antibody was added into cell lysates and enriched RNAs were isolated. Then enriched DLEU1 was analyzed by PCR. f DLEU1 co-localized with SMARCA1 in HCT8 cells as shown by RNA FISH. Green, DLEU1; Red, SMARCA1; Blue, DAPI. Scale bar, 10 μm. g the region of nt 1~ 400 in DLEU1 was important for the interaction with SMARCA1. h DLEU1 (nt 1~ 400) associated with SMARCA1 directly as shown by RNA EMSA assays. i The region of nt 700~ 1050 is indispensable for the function of DLEU1 in colorectal cancer. Overexpression of DLEU1 with deletion of nt 1~ 400 cannot promoted proliferation and metastasis in CC. *** P
    Figure Legend Snippet: DLEU1 interacts with SMARCA1 in CRC cells. a The expression of DLEU1 in cytoplasm and nucleus of HCT8 cells was measured by qRT-PCR. U6 serves as a nuclear control. GAPDH serves as a cytoplasmic control. b SMARCA1 was a potential interactive candidate of DLEU1. Biotin-labeled DLEU1 and intron control were incubated with HCT8 cell lysates, and the enriched products were eluted and separated by SDS-PAGE electrophoresis and silver staining. The differential band appearing in DLEU1 lane was analyzed by mass spectrum. c DLEU1 associated with SMARCA1 as shown by RNA pulldown and Western blot. Biotin-labeled DLEU1 and intron control were added into HCT8 cell lysates, and pulldown assays were performed. d DLEU1 was enriched by SMARCA1 in HCT8 and SW480 cell lysates. e SMARCA1 enriched DLEU1 in HCT8 cell lysates. SMARCA1 antibody was added into cell lysates and enriched RNAs were isolated. Then enriched DLEU1 was analyzed by PCR. f DLEU1 co-localized with SMARCA1 in HCT8 cells as shown by RNA FISH. Green, DLEU1; Red, SMARCA1; Blue, DAPI. Scale bar, 10 μm. g the region of nt 1~ 400 in DLEU1 was important for the interaction with SMARCA1. h DLEU1 (nt 1~ 400) associated with SMARCA1 directly as shown by RNA EMSA assays. i The region of nt 700~ 1050 is indispensable for the function of DLEU1 in colorectal cancer. Overexpression of DLEU1 with deletion of nt 1~ 400 cannot promoted proliferation and metastasis in CC. *** P

    Techniques Used: Expressing, Quantitative RT-PCR, Labeling, Incubation, SDS Page, Electrophoresis, Silver Staining, Western Blot, Isolation, Polymerase Chain Reaction, Fluorescence In Situ Hybridization, Over Expression

    34) Product Images from "lncRNA SNHG6 regulates EZH2 expression by sponging miR-26a/b and miR-214 in colorectal cancer"

    Article Title: lncRNA SNHG6 regulates EZH2 expression by sponging miR-26a/b and miR-214 in colorectal cancer

    Journal: Journal of Hematology & Oncology

    doi: 10.1186/s13045-018-0690-5

    DNA copy number gains and SP1 activation induce high SNHG6 expression in CRC. a GSEA results were plotted to visualize the correlation between the expression of SNHG6 and genes in adjacent chromosomal regions (CHR8Q11, CHR8Q12, CHR8Q13, CHR8Q21, CHR8Q22, CHR8Q23, and CHR8Q24). b SNHG6 genomic copy numbers in 30 CRC tissue samples. c SNHG6 genomic copy numbers in CRC cell lines and the normal colorectal epithelial cell line FHC. d Analysis of SP1 ChIP-seq, H3K4me3 ChIP-seq, and DnaseI-seq data of HCT-116 cells in the SNHG6 locus. e SNHG6 expression was detected by qRT-PCR in HCT-116 and HCT-8 cells transfected with the SP1 siRNAs or SP1 overexpression vector. f The correlation between the SP1 and SNHG6 expression levels were analyzed in 30 paired colorectal cancer samples ( n = 30, r = 0.431, P = 0.017). g Luciferase reporter assays were used to determine the SP1 binding sites on the SNHG6 promoter region. h ChIP assays were performed to detect SP1 occupancy in the SNHG6 promoter region. ** P
    Figure Legend Snippet: DNA copy number gains and SP1 activation induce high SNHG6 expression in CRC. a GSEA results were plotted to visualize the correlation between the expression of SNHG6 and genes in adjacent chromosomal regions (CHR8Q11, CHR8Q12, CHR8Q13, CHR8Q21, CHR8Q22, CHR8Q23, and CHR8Q24). b SNHG6 genomic copy numbers in 30 CRC tissue samples. c SNHG6 genomic copy numbers in CRC cell lines and the normal colorectal epithelial cell line FHC. d Analysis of SP1 ChIP-seq, H3K4me3 ChIP-seq, and DnaseI-seq data of HCT-116 cells in the SNHG6 locus. e SNHG6 expression was detected by qRT-PCR in HCT-116 and HCT-8 cells transfected with the SP1 siRNAs or SP1 overexpression vector. f The correlation between the SP1 and SNHG6 expression levels were analyzed in 30 paired colorectal cancer samples ( n = 30, r = 0.431, P = 0.017). g Luciferase reporter assays were used to determine the SP1 binding sites on the SNHG6 promoter region. h ChIP assays were performed to detect SP1 occupancy in the SNHG6 promoter region. ** P

    Techniques Used: Activation Assay, Expressing, Chromatin Immunoprecipitation, Quantitative RT-PCR, Transfection, Over Expression, Plasmid Preparation, Luciferase, Binding Assay

    35) Product Images from "lncRNA SNHG6 regulates EZH2 expression by sponging miR-26a/b and miR-214 in colorectal cancer"

    Article Title: lncRNA SNHG6 regulates EZH2 expression by sponging miR-26a/b and miR-214 in colorectal cancer

    Journal: Journal of Hematology & Oncology

    doi: 10.1186/s13045-018-0690-5

    SNHG6 expression is upregulated in colorectal cancer and high SNHG6 expression predicts poor prognosis. a Hierarchical cluster heat map of aberrantly expressed lncRNAs in CRC generated from RNA sequencing data from the TCGA database. Red in the heat map denotes upregulation, while blue denotes downregulation. The red line indicates SNHG6. b Expression of SNHG6 in TCGA, GSE8671, and GSE9348 CRC cohorts. c qRT-PCR analysis of SNHG6 expression in 80 pairs of colorectal cancer and corresponding normal tissues. d SNHG6 expression in CRC cell lines (DLD-1, HCT-116, HT-29, SW-620, HCT-8, and SW-480) compared with normal colorectal epithelial cell line FHC detected by qRT-PCR. e Gene set enrichment analysis results of were plotted to visualize the correlation between the expression of SNHG6 and genes dysregulated in colorectal cancer (GRADE_COLON_AND_RECTAL_CANCER_DN and GRADE_COLON_AND_RECTAL_CANCER_UP). f Kaplan–Meier survival analysis of CRC patients’ overall survival based on their SNHG6 expression, a tissue sample whose threshold cycle (CT) value of SNHG6 minus CT value of GAPDH less than − 5.56 was defined as high SNHG6 expression ( n = 120, P = 0.002). g Kaplan–Meier survival analysis of CRC patients’ disease-free survival based on their SNHG6 expression, a tissue sample whose threshold cycle (CT) value of SNHG6 minus CT value of GAPDH less than − 5.56 was defined as high SNHG6 expression ( n = 120, P = 0.036). ** P
    Figure Legend Snippet: SNHG6 expression is upregulated in colorectal cancer and high SNHG6 expression predicts poor prognosis. a Hierarchical cluster heat map of aberrantly expressed lncRNAs in CRC generated from RNA sequencing data from the TCGA database. Red in the heat map denotes upregulation, while blue denotes downregulation. The red line indicates SNHG6. b Expression of SNHG6 in TCGA, GSE8671, and GSE9348 CRC cohorts. c qRT-PCR analysis of SNHG6 expression in 80 pairs of colorectal cancer and corresponding normal tissues. d SNHG6 expression in CRC cell lines (DLD-1, HCT-116, HT-29, SW-620, HCT-8, and SW-480) compared with normal colorectal epithelial cell line FHC detected by qRT-PCR. e Gene set enrichment analysis results of were plotted to visualize the correlation between the expression of SNHG6 and genes dysregulated in colorectal cancer (GRADE_COLON_AND_RECTAL_CANCER_DN and GRADE_COLON_AND_RECTAL_CANCER_UP). f Kaplan–Meier survival analysis of CRC patients’ overall survival based on their SNHG6 expression, a tissue sample whose threshold cycle (CT) value of SNHG6 minus CT value of GAPDH less than − 5.56 was defined as high SNHG6 expression ( n = 120, P = 0.002). g Kaplan–Meier survival analysis of CRC patients’ disease-free survival based on their SNHG6 expression, a tissue sample whose threshold cycle (CT) value of SNHG6 minus CT value of GAPDH less than − 5.56 was defined as high SNHG6 expression ( n = 120, P = 0.036). ** P

    Techniques Used: Expressing, Generated, RNA Sequencing Assay, Quantitative RT-PCR

    DNA copy number gains and SP1 activation induce high SNHG6 expression in CRC. a GSEA results were plotted to visualize the correlation between the expression of SNHG6 and genes in adjacent chromosomal regions (CHR8Q11, CHR8Q12, CHR8Q13, CHR8Q21, CHR8Q22, CHR8Q23, and CHR8Q24). b SNHG6 genomic copy numbers in 30 CRC tissue samples. c SNHG6 genomic copy numbers in CRC cell lines and the normal colorectal epithelial cell line FHC. d Analysis of SP1 ChIP-seq, H3K4me3 ChIP-seq, and DnaseI-seq data of HCT-116 cells in the SNHG6 locus. e SNHG6 expression was detected by qRT-PCR in HCT-116 and HCT-8 cells transfected with the SP1 siRNAs or SP1 overexpression vector. f The correlation between the SP1 and SNHG6 expression levels were analyzed in 30 paired colorectal cancer samples ( n = 30, r = 0.431, P = 0.017). g Luciferase reporter assays were used to determine the SP1 binding sites on the SNHG6 promoter region. h ChIP assays were performed to detect SP1 occupancy in the SNHG6 promoter region. ** P
    Figure Legend Snippet: DNA copy number gains and SP1 activation induce high SNHG6 expression in CRC. a GSEA results were plotted to visualize the correlation between the expression of SNHG6 and genes in adjacent chromosomal regions (CHR8Q11, CHR8Q12, CHR8Q13, CHR8Q21, CHR8Q22, CHR8Q23, and CHR8Q24). b SNHG6 genomic copy numbers in 30 CRC tissue samples. c SNHG6 genomic copy numbers in CRC cell lines and the normal colorectal epithelial cell line FHC. d Analysis of SP1 ChIP-seq, H3K4me3 ChIP-seq, and DnaseI-seq data of HCT-116 cells in the SNHG6 locus. e SNHG6 expression was detected by qRT-PCR in HCT-116 and HCT-8 cells transfected with the SP1 siRNAs or SP1 overexpression vector. f The correlation between the SP1 and SNHG6 expression levels were analyzed in 30 paired colorectal cancer samples ( n = 30, r = 0.431, P = 0.017). g Luciferase reporter assays were used to determine the SP1 binding sites on the SNHG6 promoter region. h ChIP assays were performed to detect SP1 occupancy in the SNHG6 promoter region. ** P

    Techniques Used: Activation Assay, Expressing, Chromatin Immunoprecipitation, Quantitative RT-PCR, Transfection, Over Expression, Plasmid Preparation, Luciferase, Binding Assay

    36) Product Images from "Ficus dubia Latex Extract Induces Cell Cycle Arrest and Apoptosis by Regulating the NF-κB Pathway in Inflammatory Human Colorectal Cancer Cell Lines"

    Article Title: Ficus dubia Latex Extract Induces Cell Cycle Arrest and Apoptosis by Regulating the NF-κB Pathway in Inflammatory Human Colorectal Cancer Cell Lines

    Journal: Cancers

    doi: 10.3390/cancers14112665

    The effects of FDLE on NF-κB-mediated expression of apoptotic proteins and activation of caspase enzymes in non-inflammatory and inflammatory HCT-116 and HT-29 colorectal cancer cell lines by Western blot. ( A , B ) The expression of apoptotic proteins and the activation of caspase enzymes in non-inflammation HCT-116 and HT-29 cells untreated and treated with three doses (50, 100 and 200 µg/mL) of FDLE or BEZ235 for 48 h. ( C , D ) The expression of apoptotic proteins and the activation of caspase enzymes in inflammation HCT-116 and HT-29 cells untreated and treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h after adding cytokines (TNF-α, IFN-γ and LPS each 10 ng/mL) for 2 h. Densitometric quantification was performed by image software. The results were represented as mean ± SD of three independent experiments, * p
    Figure Legend Snippet: The effects of FDLE on NF-κB-mediated expression of apoptotic proteins and activation of caspase enzymes in non-inflammatory and inflammatory HCT-116 and HT-29 colorectal cancer cell lines by Western blot. ( A , B ) The expression of apoptotic proteins and the activation of caspase enzymes in non-inflammation HCT-116 and HT-29 cells untreated and treated with three doses (50, 100 and 200 µg/mL) of FDLE or BEZ235 for 48 h. ( C , D ) The expression of apoptotic proteins and the activation of caspase enzymes in inflammation HCT-116 and HT-29 cells untreated and treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h after adding cytokines (TNF-α, IFN-γ and LPS each 10 ng/mL) for 2 h. Densitometric quantification was performed by image software. The results were represented as mean ± SD of three independent experiments, * p

    Techniques Used: Expressing, Activation Assay, Western Blot, Software

    The effects of FDLE on apoptosis induction in non-inflammatory and inflammatory HCT-116 and HT-29 colorectal cancer cell lines. Apoptosis includes early apoptosis (LR) and late apoptosis (UR). ( A – C ) Apoptosis rate of non-inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h. ( D – F ) Apoptosis rate of inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h after adding a mixture of cytokines (TNF-α, IFN-γ and LPS each 10 ng/mL) for 2 h. The results were represented as mean ± SD of three independent experiments, * p
    Figure Legend Snippet: The effects of FDLE on apoptosis induction in non-inflammatory and inflammatory HCT-116 and HT-29 colorectal cancer cell lines. Apoptosis includes early apoptosis (LR) and late apoptosis (UR). ( A – C ) Apoptosis rate of non-inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h. ( D – F ) Apoptosis rate of inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h after adding a mixture of cytokines (TNF-α, IFN-γ and LPS each 10 ng/mL) for 2 h. The results were represented as mean ± SD of three independent experiments, * p

    Techniques Used:

    The different anti-proliferative mechanisms of FDLE on HCT-116 and HT-29 human colorectal cancer cell lines in non-inflammatory and inflammatory conditions.
    Figure Legend Snippet: The different anti-proliferative mechanisms of FDLE on HCT-116 and HT-29 human colorectal cancer cell lines in non-inflammatory and inflammatory conditions.

    Techniques Used:

    The effects of FDLE on NF-κB-mediated cell cycle proteins in non-inflammatory and inflammatory HCT-116 and HT-29 colorectal cancer cell line. ( A , B ) Western blot analysis of NF-κB-mediated proteins related to cell cycle in non-inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h. ( C , D ) Western blot analysis of NF-κB-mediated proteins related to cell cycle in inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h after adding a mixture of cytokines (TNF-α, IFN-γ and LPS each 10 ng/mL) for 2 h. Densitometric quantification was performed by image software. The results were represented as mean ± SD of three independent experiments, * p
    Figure Legend Snippet: The effects of FDLE on NF-κB-mediated cell cycle proteins in non-inflammatory and inflammatory HCT-116 and HT-29 colorectal cancer cell line. ( A , B ) Western blot analysis of NF-κB-mediated proteins related to cell cycle in non-inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h. ( C , D ) Western blot analysis of NF-κB-mediated proteins related to cell cycle in inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h after adding a mixture of cytokines (TNF-α, IFN-γ and LPS each 10 ng/mL) for 2 h. Densitometric quantification was performed by image software. The results were represented as mean ± SD of three independent experiments, * p

    Techniques Used: Western Blot, Software

    The effects of FDLE on cell cycle arrest in non-inflammatory and inflammatory HCT-116 and HT-29 colorectal cancer cells by flow cytometry. ( A – C ) Representative cell cycle distribution of non-inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h. ( D – F ) Representative cell cycle distribution of inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h after adding a mixture of cytokines (TNF-α, IFN-γ and LPS each 10 ng/mL) for 2 h. The results were represented as mean ± SD of three independent experiments, * p
    Figure Legend Snippet: The effects of FDLE on cell cycle arrest in non-inflammatory and inflammatory HCT-116 and HT-29 colorectal cancer cells by flow cytometry. ( A – C ) Representative cell cycle distribution of non-inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h. ( D – F ) Representative cell cycle distribution of inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h after adding a mixture of cytokines (TNF-α, IFN-γ and LPS each 10 ng/mL) for 2 h. The results were represented as mean ± SD of three independent experiments, * p

    Techniques Used: Flow Cytometry

    The effects of FDLE on growth inhibition in non-inflammatory and inflammatory HCT-116 and HT-29 colorectal cancer cells by colony formation assay. ( A – C ) Colony number of non-inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h. ( D – F ) Colony number of inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h after adding a mixture of cytokines (TNF-α, IFN-γ and LPS each 10 ng/mL) for 2 h. The cells were washed by PBS and cultured for 7 days. The results were represented as mean ± SD of three independent experiments, * p
    Figure Legend Snippet: The effects of FDLE on growth inhibition in non-inflammatory and inflammatory HCT-116 and HT-29 colorectal cancer cells by colony formation assay. ( A – C ) Colony number of non-inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h. ( D – F ) Colony number of inflammatory HCT-116 and HT-29 cells treated with three doses (50, 100 and 200 µg/mL) of FDLE for 48 h after adding a mixture of cytokines (TNF-α, IFN-γ and LPS each 10 ng/mL) for 2 h. The cells were washed by PBS and cultured for 7 days. The results were represented as mean ± SD of three independent experiments, * p

    Techniques Used: Inhibition, Colony Assay, Cell Culture

    The effects of FDLE on cell viability of colorectal cancer cell lines (HCT-116 and HT-29) and normal mouse embryonic fibroblast cell (NIH3T3) in non-inflammatory and inflammatory states by MTT. The cells were treated with various concentrations of FDLE for 24, 48 and 72 h. The cell viability was calculated comparing to untreated control cells after 24, 48 and 72 h of incubation. ( A – C ) The cell viability of HCT-116, HT-29 and NIH3T3 cells in non-inflammatory state. ( D – F ) The cell viability of HCT-116, HT-29 and NIH3T3 cells in inflammatory state. The results were represented as mean ± SD of three independent experiments, * p
    Figure Legend Snippet: The effects of FDLE on cell viability of colorectal cancer cell lines (HCT-116 and HT-29) and normal mouse embryonic fibroblast cell (NIH3T3) in non-inflammatory and inflammatory states by MTT. The cells were treated with various concentrations of FDLE for 24, 48 and 72 h. The cell viability was calculated comparing to untreated control cells after 24, 48 and 72 h of incubation. ( A – C ) The cell viability of HCT-116, HT-29 and NIH3T3 cells in non-inflammatory state. ( D – F ) The cell viability of HCT-116, HT-29 and NIH3T3 cells in inflammatory state. The results were represented as mean ± SD of three independent experiments, * p

    Techniques Used: MTT Assay, Incubation

    37) Product Images from "Silencing or inhibition of H3K79 methyltransferase DOT1L induces cell cycle arrest by epigenetically modulating c-Myc expression in colorectal cancer"

    Article Title: Silencing or inhibition of H3K79 methyltransferase DOT1L induces cell cycle arrest by epigenetically modulating c-Myc expression in colorectal cancer

    Journal: Clinical Epigenetics

    doi: 10.1186/s13148-019-0778-y

    DOT1L silencing or inhibition demethylates H3K79 and suppresses transcription of c-Myc. a The Pearson correlation between DOT1L and c-Myc expression in patients with colorectal cancer in the TCGA COAd datasheet from the GEPIA. b Relative mRNA expression of c-Myc in patients with colorectal cancer in the TCGA COAd datasheet from the GEPIA. c Protein expression of c-Myc in SW480 and HCT116 cells after DOT1L knockdown or inhibition. Gray ratio of each blot was analyzed by using the Image J software and protein/GAPDH ratio was shown. d mRNA expression of c-Myc, CDK2, Cyclin A2 in SW480, and HCT116 cells after DOT1L knockdown or inhibition. e H3K9 methylation (m1/2/3) was detected by using Western blot in SW480 and HCT116 cells after DOT1L knockdown or inhibition. Gray ration of each blot was analyzed by using the Image J software and protein/H3 ratio was shown. f–h ChIP assay was performed to detect the binding region of H3K79me2 on the promoter of c-Myc in SW480 and HCT116 cells after DOT1L knockdown or inhibition. i , j IHC staining of c-Myc and H3K79me1/2/3 in xenografts obtained from subcutaneous injecting SW480 and HCT116 cells after DOT1L knockdown or inhibition within mice. Signal-positive rate was analyzed by using the IHC profiler in the Image J software
    Figure Legend Snippet: DOT1L silencing or inhibition demethylates H3K79 and suppresses transcription of c-Myc. a The Pearson correlation between DOT1L and c-Myc expression in patients with colorectal cancer in the TCGA COAd datasheet from the GEPIA. b Relative mRNA expression of c-Myc in patients with colorectal cancer in the TCGA COAd datasheet from the GEPIA. c Protein expression of c-Myc in SW480 and HCT116 cells after DOT1L knockdown or inhibition. Gray ratio of each blot was analyzed by using the Image J software and protein/GAPDH ratio was shown. d mRNA expression of c-Myc, CDK2, Cyclin A2 in SW480, and HCT116 cells after DOT1L knockdown or inhibition. e H3K9 methylation (m1/2/3) was detected by using Western blot in SW480 and HCT116 cells after DOT1L knockdown or inhibition. Gray ration of each blot was analyzed by using the Image J software and protein/H3 ratio was shown. f–h ChIP assay was performed to detect the binding region of H3K79me2 on the promoter of c-Myc in SW480 and HCT116 cells after DOT1L knockdown or inhibition. i , j IHC staining of c-Myc and H3K79me1/2/3 in xenografts obtained from subcutaneous injecting SW480 and HCT116 cells after DOT1L knockdown or inhibition within mice. Signal-positive rate was analyzed by using the IHC profiler in the Image J software

    Techniques Used: Inhibition, Expressing, Software, Methylation, Western Blot, Chromatin Immunoprecipitation, Binding Assay, Immunohistochemistry, Staining, Mouse Assay

    DOT1L is highly expressed in colorectal cancer than other cancer types. a The relative mRNA expression of DOT1L in multiple cancer types in Bittner Multi-cancer datasheet from the Oncomine. b DOT1L DNA copy number in multiple cancer types in Beroukhim multi-cancer datasheet from the Oncomine
    Figure Legend Snippet: DOT1L is highly expressed in colorectal cancer than other cancer types. a The relative mRNA expression of DOT1L in multiple cancer types in Bittner Multi-cancer datasheet from the Oncomine. b DOT1L DNA copy number in multiple cancer types in Beroukhim multi-cancer datasheet from the Oncomine

    Techniques Used: Expressing

    DOT1L is upregulated in colorectal cancer, especially in colorectal carcinoma. a Immunohistochemical (IHC) staining analysis of DOT1L expression in colorectal cancer ( N = 45) tissues and normal tissues ( N = 46). b Relative mRNA expression of DOT1L in normal colon or CRC tissues in Hong colorectal datasheet from the Oncomine. c Relative mRNA expression of DOT1L in normal rectum or rectum adenocarcinoma tissues in Gaedcke colorectal datasheet from the Oncomine. d Relative mRNA expression of DOT1L in normal colon or COAD tissues in Skrzypczak colorectal 2 datasheet from the Oncomine. e DOT1L promoter methylation in COAD or normal tissues from DiseaseMeth v.2. f DOT1L promoter methylation in READ or normal tissues from DiseaseMeth v.2. g Relative mRNA expression of DOT1L in COAD or READ tissues in Jorissen colorectal 3 datasheet from the Oncomine. h Relative mRNA expression of DOT1L in colorectal adenoma or colorectal carcinoma tissues in Skrzypczak colorectal 2 datasheet from the Oncomine
    Figure Legend Snippet: DOT1L is upregulated in colorectal cancer, especially in colorectal carcinoma. a Immunohistochemical (IHC) staining analysis of DOT1L expression in colorectal cancer ( N = 45) tissues and normal tissues ( N = 46). b Relative mRNA expression of DOT1L in normal colon or CRC tissues in Hong colorectal datasheet from the Oncomine. c Relative mRNA expression of DOT1L in normal rectum or rectum adenocarcinoma tissues in Gaedcke colorectal datasheet from the Oncomine. d Relative mRNA expression of DOT1L in normal colon or COAD tissues in Skrzypczak colorectal 2 datasheet from the Oncomine. e DOT1L promoter methylation in COAD or normal tissues from DiseaseMeth v.2. f DOT1L promoter methylation in READ or normal tissues from DiseaseMeth v.2. g Relative mRNA expression of DOT1L in COAD or READ tissues in Jorissen colorectal 3 datasheet from the Oncomine. h Relative mRNA expression of DOT1L in colorectal adenoma or colorectal carcinoma tissues in Skrzypczak colorectal 2 datasheet from the Oncomine

    Techniques Used: Immunohistochemistry, Staining, Expressing, Methylation

    DOT1L silencing or inhibition blocks cell proliferation of CRC cells in vitro. a Relative mRNA expression of DOT1L detected by using qRT-PCR in SW480 and HCT116 colorectal cancer cell lines after DOT1L knockdown. shGFP vectors were used as control. b Protein expression of DOT1L detected by using Western blot in SW480 and HCT116 cells after DOT1L knockdown. Gray ratio of each blot was analyzed by using the Image J software and DOT1L/GAPDH ratio was shown. c , d Cell growth curve was determined by using MTT assay in SW480 and HCT116 cells after DOT1L knockdown or treatment with its specific inhibitor EPZ004777 with different concentrations for 1/3/5/7 days. e, f Cell proliferation was detected by using BrdU immunofluorescence in SW480 and HCT116 cells after DOT1L knockdown or inhibited by using EPZ004777 for 48 h (30 μM in SW480 and 50 μM in HCT116)
    Figure Legend Snippet: DOT1L silencing or inhibition blocks cell proliferation of CRC cells in vitro. a Relative mRNA expression of DOT1L detected by using qRT-PCR in SW480 and HCT116 colorectal cancer cell lines after DOT1L knockdown. shGFP vectors were used as control. b Protein expression of DOT1L detected by using Western blot in SW480 and HCT116 cells after DOT1L knockdown. Gray ratio of each blot was analyzed by using the Image J software and DOT1L/GAPDH ratio was shown. c , d Cell growth curve was determined by using MTT assay in SW480 and HCT116 cells after DOT1L knockdown or treatment with its specific inhibitor EPZ004777 with different concentrations for 1/3/5/7 days. e, f Cell proliferation was detected by using BrdU immunofluorescence in SW480 and HCT116 cells after DOT1L knockdown or inhibited by using EPZ004777 for 48 h (30 μM in SW480 and 50 μM in HCT116)

    Techniques Used: Inhibition, In Vitro, Expressing, Quantitative RT-PCR, Western Blot, Software, MTT Assay, Immunofluorescence

    38) Product Images from "Quinacrine-Mediated Inhibition of Nrf2 Reverses Hypoxia-Induced 5-Fluorouracil Resistance in Colorectal Cancer"

    Article Title: Quinacrine-Mediated Inhibition of Nrf2 Reverses Hypoxia-Induced 5-Fluorouracil Resistance in Colorectal Cancer

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20184366

    QC decreases Nrf2 protein stability in CRC cells. ( A,B ) Effects of QC on Nrf2 mRNA expression ( A ) and Nrf2 mRNA stability ( B ) in HCT116 cells (left) and RKO cells (right). ( C,D ) Effect of QC on Nrf2 protein stability in HCT116 cells ( C ) and RKO cells ( D ) under hypoxic conditions. Nrf2 protein levels were quantified using Image J software; band intensities were normalized to those of β-actin (band intensity at t 0 was defined as 100%). Representative images of Nrf2 and β-actin were detected by immunoblot. Representative images of immunoblots for each protein were obtained using the same sample on different gels after a single experiment.
    Figure Legend Snippet: QC decreases Nrf2 protein stability in CRC cells. ( A,B ) Effects of QC on Nrf2 mRNA expression ( A ) and Nrf2 mRNA stability ( B ) in HCT116 cells (left) and RKO cells (right). ( C,D ) Effect of QC on Nrf2 protein stability in HCT116 cells ( C ) and RKO cells ( D ) under hypoxic conditions. Nrf2 protein levels were quantified using Image J software; band intensities were normalized to those of β-actin (band intensity at t 0 was defined as 100%). Representative images of Nrf2 and β-actin were detected by immunoblot. Representative images of immunoblots for each protein were obtained using the same sample on different gels after a single experiment.

    Techniques Used: Expressing, Software, Western Blot

    QC sensitizes CRC cells to 5-FU in hypoxia by inhibiting nuclear factor (erythroid-derived 2)-like 2 (Nrf2). ( A ) Scheme of the treatment and sampling procedure. ( B,C ) Effect of the QC/5-FU combination on Nrf2 expression in HCT116 ( B ) and RKO ( C ) cells under hypoxic conditions. ( D ) Effect of the QC/5-FU combination on Nrf2 expression in HT-29, DLD1, SW480, SW620, HCT15, and Colo205 cells under hypoxic conditions. Representative images of Nrf2 and β-actin were detected by immunoblot. Representative images of immunoblots for each protein were obtained using the same sample on different gels after a single experiment. ( E,F ) Effect of Nrf2 overexpression on DNA damage induced in HCT116 ( E ) and RKO ( F ) cells by the QC/5-FU combination under hypoxic conditions. γ-H2AX (green) staining in HCT116 ( E ) and RKO ( F ) cells following treatment (left), and the relative percentage of foci-positive cells (right). Data are presented as means ± SD (** p
    Figure Legend Snippet: QC sensitizes CRC cells to 5-FU in hypoxia by inhibiting nuclear factor (erythroid-derived 2)-like 2 (Nrf2). ( A ) Scheme of the treatment and sampling procedure. ( B,C ) Effect of the QC/5-FU combination on Nrf2 expression in HCT116 ( B ) and RKO ( C ) cells under hypoxic conditions. ( D ) Effect of the QC/5-FU combination on Nrf2 expression in HT-29, DLD1, SW480, SW620, HCT15, and Colo205 cells under hypoxic conditions. Representative images of Nrf2 and β-actin were detected by immunoblot. Representative images of immunoblots for each protein were obtained using the same sample on different gels after a single experiment. ( E,F ) Effect of Nrf2 overexpression on DNA damage induced in HCT116 ( E ) and RKO ( F ) cells by the QC/5-FU combination under hypoxic conditions. γ-H2AX (green) staining in HCT116 ( E ) and RKO ( F ) cells following treatment (left), and the relative percentage of foci-positive cells (right). Data are presented as means ± SD (** p

    Techniques Used: Derivative Assay, Sampling, Expressing, Western Blot, Over Expression, Staining

    Quinacrine (QC) sensitizes colorectal cancer (CRC) cells to 5-fluorouracil (5-FU) treatment under hypoxic conditions. ( A ) ATP-Glo assay of the QC or 5-FU treatments in HCT116, HT29, DLD1, RKO, SW620, and Colo205 cells. ( B ) Summary of IC 50 values and associated 95% confidence intervals (CI) for QC, 5-FU, and combined QC and 5-FU treatment in all tested CRC cell lines. ( C ) Clonogenic survival assay for the QC/5-FU combination and single-agent treatments in HCT116, HT29, DLD1, RKO, SW620, and Colo205 cells. Data are presented as means ± SD (* p
    Figure Legend Snippet: Quinacrine (QC) sensitizes colorectal cancer (CRC) cells to 5-fluorouracil (5-FU) treatment under hypoxic conditions. ( A ) ATP-Glo assay of the QC or 5-FU treatments in HCT116, HT29, DLD1, RKO, SW620, and Colo205 cells. ( B ) Summary of IC 50 values and associated 95% confidence intervals (CI) for QC, 5-FU, and combined QC and 5-FU treatment in all tested CRC cell lines. ( C ) Clonogenic survival assay for the QC/5-FU combination and single-agent treatments in HCT116, HT29, DLD1, RKO, SW620, and Colo205 cells. Data are presented as means ± SD (* p

    Techniques Used: Glo Assay, Clonogenic Cell Survival Assay

    JNK1 activation is required for the QC-induced degradation of Nrf2 protein. ( A,B ) Effect of QC on the expression of Nrf2, Keap1, Cul3, p-p38, p38, pERK1/2 (phospho extracellular signal-regulated kinases), ERK1/2, p-JNK, JNK, and β-actin in HCT116 cells ( A ) and RKO cells ( B ) under normoxic and hypoxic conditions. ( C,D ) Effect of QC on the proteasome-mediated degradation of Nrf2 in HCT116 cells ( C ) and RKO cells ( D ) under normoxic and hypoxic conditions. ( E,F ) Effect of QC on Keap1/Cul3-dependent degradation of Nrf2 in HCT116 cells ( E ) and RKO cells ( F ) under normoxic and hypoxic conditions. ( G,H ) Effect of JNK1 activation on QC-induced inhibition of Nrf2 in HCT116 cells ( G ) and RKO cells ( H ) under normoxic and hypoxic conditions. ( I,J ) Effect of QC on the interaction between Nrf2 and Keap1 in HCT116 cells ( I ) and RKO cells ( J ) under hypoxic conditions. Representative images of Nrf2, Keap1, Cul3, p-p38, p38, pERK1/2, ERK1/2, pJNK, JNK, and β-actin were detected by immunoblot. Representative images of immunoblots for each protein were obtained using the same sample on different gels after a single experiment.
    Figure Legend Snippet: JNK1 activation is required for the QC-induced degradation of Nrf2 protein. ( A,B ) Effect of QC on the expression of Nrf2, Keap1, Cul3, p-p38, p38, pERK1/2 (phospho extracellular signal-regulated kinases), ERK1/2, p-JNK, JNK, and β-actin in HCT116 cells ( A ) and RKO cells ( B ) under normoxic and hypoxic conditions. ( C,D ) Effect of QC on the proteasome-mediated degradation of Nrf2 in HCT116 cells ( C ) and RKO cells ( D ) under normoxic and hypoxic conditions. ( E,F ) Effect of QC on Keap1/Cul3-dependent degradation of Nrf2 in HCT116 cells ( E ) and RKO cells ( F ) under normoxic and hypoxic conditions. ( G,H ) Effect of JNK1 activation on QC-induced inhibition of Nrf2 in HCT116 cells ( G ) and RKO cells ( H ) under normoxic and hypoxic conditions. ( I,J ) Effect of QC on the interaction between Nrf2 and Keap1 in HCT116 cells ( I ) and RKO cells ( J ) under hypoxic conditions. Representative images of Nrf2, Keap1, Cul3, p-p38, p38, pERK1/2, ERK1/2, pJNK, JNK, and β-actin were detected by immunoblot. Representative images of immunoblots for each protein were obtained using the same sample on different gels after a single experiment.

    Techniques Used: Activation Assay, Expressing, Inhibition, Western Blot

    39) Product Images from "Bioactive Vitamin D Attenuates MED28-Mediated Cell Growth and Epithelial–Mesenchymal Transition in Human Colorectal Cancer Cells"

    Article Title: Bioactive Vitamin D Attenuates MED28-Mediated Cell Growth and Epithelial–Mesenchymal Transition in Human Colorectal Cancer Cells

    Journal: BioMed Research International

    doi: 10.1155/2022/2268818

    Calcitriol suppressed the upregulatory effect of MED28 on cell growth and epithelial-mesenchymal transition in SW480 human colorectal cancer cells. SW480 cells were treated with calcitriol (100 nM) for 48 h (a), undergone MED28-specific siRNA (siMED28) for 72 h (b), or MED28 overexpression plasmid (Flag-MED28) for 48 h (c), with respective controls, and subjected to Western blotting. The ratios below the representative images (left panels) indicate the relative expression of the specific proteins with respect to those of 0 nM (a), siControl (b), or pcDNA3 (c) after normalization with the expression of the corresponding loading controls, β -actin, α -tubulin, GAPDH, or vinculin. Densitometric data (right panels) are expressed as means ± standard deviation, n = 3; ∗∗ p
    Figure Legend Snippet: Calcitriol suppressed the upregulatory effect of MED28 on cell growth and epithelial-mesenchymal transition in SW480 human colorectal cancer cells. SW480 cells were treated with calcitriol (100 nM) for 48 h (a), undergone MED28-specific siRNA (siMED28) for 72 h (b), or MED28 overexpression plasmid (Flag-MED28) for 48 h (c), with respective controls, and subjected to Western blotting. The ratios below the representative images (left panels) indicate the relative expression of the specific proteins with respect to those of 0 nM (a), siControl (b), or pcDNA3 (c) after normalization with the expression of the corresponding loading controls, β -actin, α -tubulin, GAPDH, or vinculin. Densitometric data (right panels) are expressed as means ± standard deviation, n = 3; ∗∗ p

    Techniques Used: Over Expression, Plasmid Preparation, Western Blot, Expressing, Standard Deviation

    Calcitriol suppresses Wnt/ β -catenin signaling, cell growth, and epithelial-mesenchymal transition, partially through downregulating MED28 expression, in human colorectal cancer cells. (a) SW480 and HT29 cells were treated with calcitriol (100 nM) for 48 h and subjected to RNA extraction and quantitative real-time polymerase chain reaction as described in Materials and Methods. Data are expressed as means ± standard deviation, n = 3; ∗ p
    Figure Legend Snippet: Calcitriol suppresses Wnt/ β -catenin signaling, cell growth, and epithelial-mesenchymal transition, partially through downregulating MED28 expression, in human colorectal cancer cells. (a) SW480 and HT29 cells were treated with calcitriol (100 nM) for 48 h and subjected to RNA extraction and quantitative real-time polymerase chain reaction as described in Materials and Methods. Data are expressed as means ± standard deviation, n = 3; ∗ p

    Techniques Used: Expressing, RNA Extraction, Real-time Polymerase Chain Reaction, Standard Deviation

    Calcitriol reduced the expression of MED28, PCNA, β -catenin, c-Myc, and cyclin D1 but upregulated the expression of E-cadherin in the subcutaneous HT29 xenografts. Four-week-old male NOD/SCID mice were subcutaneously implanted with human colorectal cancer HT29 cells to their flank regions, and these animals were then randomly allotted into three groups, vehicle control (0 μ g) or calcitriol (0.5 μ g or 1 μ g), with four mice in each group. Two weeks later, the animals were intraperitoneally administered with the assigned dose, 0 μ g (vehicle), 0.5 μ g, or 1 μ g of calcitriol every other day. The animals were sacrificed after 4 weeks. (a) Representative images and densitometric quantification for the relative protein expression of the xenografts. The expression levels of E-cadherin, MED28, PCNA, β -catenin, c-Myc, and cyclin D1 in the xenografts, along with loading controls, were detected by Western blotting. The ratios below the images (left panel) indicate the relative expression of the specific proteins with respect to those of 0 μ g after normalization with the expression of the corresponding loading controls, β -actin or α -tubulin. Densitometric data (right panel) are expressed as means ± standard deviation, n = 3; ∗ p
    Figure Legend Snippet: Calcitriol reduced the expression of MED28, PCNA, β -catenin, c-Myc, and cyclin D1 but upregulated the expression of E-cadherin in the subcutaneous HT29 xenografts. Four-week-old male NOD/SCID mice were subcutaneously implanted with human colorectal cancer HT29 cells to their flank regions, and these animals were then randomly allotted into three groups, vehicle control (0 μ g) or calcitriol (0.5 μ g or 1 μ g), with four mice in each group. Two weeks later, the animals were intraperitoneally administered with the assigned dose, 0 μ g (vehicle), 0.5 μ g, or 1 μ g of calcitriol every other day. The animals were sacrificed after 4 weeks. (a) Representative images and densitometric quantification for the relative protein expression of the xenografts. The expression levels of E-cadherin, MED28, PCNA, β -catenin, c-Myc, and cyclin D1 in the xenografts, along with loading controls, were detected by Western blotting. The ratios below the images (left panel) indicate the relative expression of the specific proteins with respect to those of 0 μ g after normalization with the expression of the corresponding loading controls, β -actin or α -tubulin. Densitometric data (right panel) are expressed as means ± standard deviation, n = 3; ∗ p

    Techniques Used: Expressing, Mouse Assay, Western Blot, Standard Deviation

    MED28 knockdown mimicked the effect of calcitriol on E-cadherin and Wnt/ β -catenin signaling in HT29 human colorectal cancer cells. HT29 cells were treated with calcitriol (100 nM) for 48 h (a) or undergone MED28-specific siRNA (siMED28) for 72 h (b) with respective controls and subjected to Western blotting with the antibodies indicated. The ratios below the representative images (left panels) indicate the relative expression of the specific proteins with respect to those of 0 nM (a) or siControl (b) after normalization with the expression of the corresponding loading controls, β -actin or α -tubulin. Densitometric data (right panels) are expressed as means ± standard deviation, n = 3; ∗ p
    Figure Legend Snippet: MED28 knockdown mimicked the effect of calcitriol on E-cadherin and Wnt/ β -catenin signaling in HT29 human colorectal cancer cells. HT29 cells were treated with calcitriol (100 nM) for 48 h (a) or undergone MED28-specific siRNA (siMED28) for 72 h (b) with respective controls and subjected to Western blotting with the antibodies indicated. The ratios below the representative images (left panels) indicate the relative expression of the specific proteins with respect to those of 0 nM (a) or siControl (b) after normalization with the expression of the corresponding loading controls, β -actin or α -tubulin. Densitometric data (right panels) are expressed as means ± standard deviation, n = 3; ∗ p

    Techniques Used: Western Blot, Expressing, Standard Deviation

    40) Product Images from "The long noncoding RNA SNHG1 regulates colorectal cancer cell growth through interactions with EZH2 and miR-154-5p"

    Article Title: The long noncoding RNA SNHG1 regulates colorectal cancer cell growth through interactions with EZH2 and miR-154-5p

    Journal: Molecular Cancer

    doi: 10.1186/s12943-018-0894-x

    SNHG1 acts as a sponge for miR-154-5p in the cytoplasm. a Representative FISH images indicated subcellular location of SNHG1 in HCT-116 and HCT-8 cells (red). Nuclei were stained by DAPI (blue). SNHG1 sense probe was employed as a negative control. b Relative SNHG1 expression levels in nuclear and cytosolic fractions of HCT-116 and HCT-8 cells. Nuclear controls: U6; Cytosolic controls: GAPDH. c Dual luciferase reporter assays were used to determine miRNAs that directly interacted with SNHG1. Luciferase activity is presented as relative luciferase activity normalized to activity of their respective negative control. d Dual luciferase reporter assays were conducted with wild type and mutant type (putative binding sites for miR-154-5p were mutated) luciferase reporter vectors. Right panel, sequence alignment of miR-154-5p and its predicted binding sites (green) in SNHG1. Predicted miR-154-5p target sequence (blue) in SNHG1 (Luc-SNHG1-wt) and position of mutated nucleotides (red) in SNHG1 (Luc-SNHG1-mut). e RNA immunoprecipitation with an anti-Ago2 antibody was used to assess endogenous Ago2 binding to RNA in HCT-116 cells, IgG was used as the control. SNHG1 and miR-154-5p levels were determined by qRT–PCR and presented as fold enrichment in Ago2 relative to input. RIP efficiency of Ago2 protein was detected by western blot. f RNA pull-down assays were used to examine the interaction of SNHG1 and Ago2 in HCT-116 cells. g CCK-8 assays demonstrated that SNHG1 silencing inhibited HCT-116 cell growth. MiR-154-5p down-regulation rescued growth inhibition caused by SNHG1 knockdown. h EdU assays revealed that SNHG1 overexpression promotes HCT-116 cell proliferation. Co-transfecting miR-154-5p mimics with the SNHG1 plasmid abolished the increased proliferation rates. i The correlation between miR-154-5p and SNHG1 expression analyzed in 30 paired colorectal cancer samples (n = 30, r = − 0.48, P = 0.008). Scale bar = 50 μm. * P
    Figure Legend Snippet: SNHG1 acts as a sponge for miR-154-5p in the cytoplasm. a Representative FISH images indicated subcellular location of SNHG1 in HCT-116 and HCT-8 cells (red). Nuclei were stained by DAPI (blue). SNHG1 sense probe was employed as a negative control. b Relative SNHG1 expression levels in nuclear and cytosolic fractions of HCT-116 and HCT-8 cells. Nuclear controls: U6; Cytosolic controls: GAPDH. c Dual luciferase reporter assays were used to determine miRNAs that directly interacted with SNHG1. Luciferase activity is presented as relative luciferase activity normalized to activity of their respective negative control. d Dual luciferase reporter assays were conducted with wild type and mutant type (putative binding sites for miR-154-5p were mutated) luciferase reporter vectors. Right panel, sequence alignment of miR-154-5p and its predicted binding sites (green) in SNHG1. Predicted miR-154-5p target sequence (blue) in SNHG1 (Luc-SNHG1-wt) and position of mutated nucleotides (red) in SNHG1 (Luc-SNHG1-mut). e RNA immunoprecipitation with an anti-Ago2 antibody was used to assess endogenous Ago2 binding to RNA in HCT-116 cells, IgG was used as the control. SNHG1 and miR-154-5p levels were determined by qRT–PCR and presented as fold enrichment in Ago2 relative to input. RIP efficiency of Ago2 protein was detected by western blot. f RNA pull-down assays were used to examine the interaction of SNHG1 and Ago2 in HCT-116 cells. g CCK-8 assays demonstrated that SNHG1 silencing inhibited HCT-116 cell growth. MiR-154-5p down-regulation rescued growth inhibition caused by SNHG1 knockdown. h EdU assays revealed that SNHG1 overexpression promotes HCT-116 cell proliferation. Co-transfecting miR-154-5p mimics with the SNHG1 plasmid abolished the increased proliferation rates. i The correlation between miR-154-5p and SNHG1 expression analyzed in 30 paired colorectal cancer samples (n = 30, r = − 0.48, P = 0.008). Scale bar = 50 μm. * P

    Techniques Used: Fluorescence In Situ Hybridization, Staining, Negative Control, Expressing, Luciferase, Activity Assay, Mutagenesis, Binding Assay, Sequencing, Immunoprecipitation, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Inhibition, Over Expression, Plasmid Preparation

    SNHG 1 promotes colorectal cancer progression partly by regulating KLF2 and CDKN2B expression. a Left panel, CCK-8 assays demonstrated that silence of SNHG1 inhibited cancer cell growth. KLF2 knockdown could rescue growth inhibition caused by SNHG1 knockdown in HCT-116 cells. Right panel, CCK-8 assays demonstrated that silence of SNHG1 inhibited cancer cell growth. CDKN2B (P15) knockdown could rescue growth inhibition caused by SNHG1 knockdown in HCT-116 cells. b EdU assays showed that SNHG1 knockdown inhibited cancer cell proliferation. Co-transfecting KLF2 or CDKN2B siRNAs with SNHG1 siRNAs reversed the decreased proliferation rates in HCT-116 cells. c EdU assays showed that EZH2 knockdown could inhibit proliferation promotion caused by SNHG1 overexpression in HCT-116 cells. d Immunohistochemistry analysis of EZH2, KLF2 and CDKN2B protein levels in colorectal cancer and normal tissues. e Immunohistochemistry analysis of KLF2 and CDKN2B protein levels in tumor tissues formed from SNHG1 knockdown or control cells. f Schematic of the proposed mechanism of SNHG1 in colorectal cancer cells. In the cytoplasm, SNHG1 acts as a ceRNA to sponge miR-154-5p and upregulated the expression of CCND2 (CyclinD2). In the nucleus, SNHG1 is involved in PRC2 mediated epigenetic repression of KLF2 and CDKN2B (P15). KLF2 is also an upstream regulatory factor of CDKN2B. Besides, CDKN2B is a well-studied inhibitor of CCND2. Downstream genes of SNHG1 formed a regulatory network to regulate growth of colorectal cancer. Scale bar = 50 μm. * P
    Figure Legend Snippet: SNHG 1 promotes colorectal cancer progression partly by regulating KLF2 and CDKN2B expression. a Left panel, CCK-8 assays demonstrated that silence of SNHG1 inhibited cancer cell growth. KLF2 knockdown could rescue growth inhibition caused by SNHG1 knockdown in HCT-116 cells. Right panel, CCK-8 assays demonstrated that silence of SNHG1 inhibited cancer cell growth. CDKN2B (P15) knockdown could rescue growth inhibition caused by SNHG1 knockdown in HCT-116 cells. b EdU assays showed that SNHG1 knockdown inhibited cancer cell proliferation. Co-transfecting KLF2 or CDKN2B siRNAs with SNHG1 siRNAs reversed the decreased proliferation rates in HCT-116 cells. c EdU assays showed that EZH2 knockdown could inhibit proliferation promotion caused by SNHG1 overexpression in HCT-116 cells. d Immunohistochemistry analysis of EZH2, KLF2 and CDKN2B protein levels in colorectal cancer and normal tissues. e Immunohistochemistry analysis of KLF2 and CDKN2B protein levels in tumor tissues formed from SNHG1 knockdown or control cells. f Schematic of the proposed mechanism of SNHG1 in colorectal cancer cells. In the cytoplasm, SNHG1 acts as a ceRNA to sponge miR-154-5p and upregulated the expression of CCND2 (CyclinD2). In the nucleus, SNHG1 is involved in PRC2 mediated epigenetic repression of KLF2 and CDKN2B (P15). KLF2 is also an upstream regulatory factor of CDKN2B. Besides, CDKN2B is a well-studied inhibitor of CCND2. Downstream genes of SNHG1 formed a regulatory network to regulate growth of colorectal cancer. Scale bar = 50 μm. * P

    Techniques Used: Expressing, CCK-8 Assay, Inhibition, Over Expression, Immunohistochemistry

    SNHG1 regulates expression of the miR-154-5p target gene, CCND2. a CCND2 expression was detected by qRT-PCR in SNHG1 siRNAs transfected or SNHG1 siRNAs and miR-154-5p inhibitors co-transfected HCT-116 cells. b CCND2 expression was detected by qRT-PCR in SNHG1 vector transfected or SNHG1 vector and miR-154-5p mimics co-transfected HCT-116 cells. c Western blot analyses of CCND2 expression after knockdown of SNHG1, overexpression of miR-154-5p or knockdown of SNHG1 + inhibition of miR-154-5p in HCT-116 cells. d Dual luciferase reporter assays demonstrated that miR-154-5p overexpression reduced Luc-CCND2 luciferase activity and SNHG1 overexpression abolished miR-154-5p induced reductions in luciferase activity in HCT-116 cells. e CCND2 expression was measured by western blot after silencing of endogenous SNHG1 and transfection with either SNHG1-mut vector, which contains mutations at the putative miR-154-5p binding site, or SNHG1 vector in HCT-116 cells. f The correlation between CCND2 and SNHG1 expression analyzed in 30 paired colorectal cancer samples ( n = 30, r = 0.38, P = 0.036). g EdU assays demonstrated HCT-116 cells proliferation rates after knockdown of SNHG1, knockdown of CCND2 or both knockdown of SNHG1and CCND2. h CCK-8 assays demonstrated that CCND2 knockdown could reverse growth promotion caused by SNHG1 overexpression in HCT-116 cells. i Immunohistochemistry analysis of CCND2 protein levels in tumor tissues formed from SNHG1 knockdown or control cells. j Detection of CCND2 protein levels in colorectal cancer and normal tissues by IHC. Scale bar = 50 μm. * P
    Figure Legend Snippet: SNHG1 regulates expression of the miR-154-5p target gene, CCND2. a CCND2 expression was detected by qRT-PCR in SNHG1 siRNAs transfected or SNHG1 siRNAs and miR-154-5p inhibitors co-transfected HCT-116 cells. b CCND2 expression was detected by qRT-PCR in SNHG1 vector transfected or SNHG1 vector and miR-154-5p mimics co-transfected HCT-116 cells. c Western blot analyses of CCND2 expression after knockdown of SNHG1, overexpression of miR-154-5p or knockdown of SNHG1 + inhibition of miR-154-5p in HCT-116 cells. d Dual luciferase reporter assays demonstrated that miR-154-5p overexpression reduced Luc-CCND2 luciferase activity and SNHG1 overexpression abolished miR-154-5p induced reductions in luciferase activity in HCT-116 cells. e CCND2 expression was measured by western blot after silencing of endogenous SNHG1 and transfection with either SNHG1-mut vector, which contains mutations at the putative miR-154-5p binding site, or SNHG1 vector in HCT-116 cells. f The correlation between CCND2 and SNHG1 expression analyzed in 30 paired colorectal cancer samples ( n = 30, r = 0.38, P = 0.036). g EdU assays demonstrated HCT-116 cells proliferation rates after knockdown of SNHG1, knockdown of CCND2 or both knockdown of SNHG1and CCND2. h CCK-8 assays demonstrated that CCND2 knockdown could reverse growth promotion caused by SNHG1 overexpression in HCT-116 cells. i Immunohistochemistry analysis of CCND2 protein levels in tumor tissues formed from SNHG1 knockdown or control cells. j Detection of CCND2 protein levels in colorectal cancer and normal tissues by IHC. Scale bar = 50 μm. * P

    Techniques Used: Expressing, Quantitative RT-PCR, Transfection, Plasmid Preparation, Western Blot, Over Expression, Inhibition, Luciferase, Activity Assay, Binding Assay, CCK-8 Assay, Immunohistochemistry

    SNHG1 participates in epigenetic repression of KLF2 and CDKN2B by interacting with PRC2. a GSEA showed a significant correlation between the SNHG1 and genes in PRC2 related pathway. b Scatter plot showing the expression relationship among SNHG1, EZH2, SUZ12 and EED in colorectal tumor tissues from TCGA database. The upper right squares show the Pearson correlation between each other. c RIPs experiments for EZH2, SUZ12 and EED were performed and the coprecipitated RNA was subjected to qRT-PCR for SNHG1. GAPDH was employed as a negative control. d RNA pull-down was used to examine the association of SNHG1 and EZH2. AR binding to HuR was used as a positive control. e PRC2 target genes expression was detected by qRT-PCR in SNHG1 siRNAs transfected HCT-116 cells. f CDKN2B and KLF2 expression was detected by qRT-PCR in SNHG1 vector transfected or SNHG1 vector and EZH2 siRNAs co-transfected CRC cells. g CDKN2B and KLF2 protein levels were detected by western blot in indicated conditions. h ChIP assays were performed to detect EZH2 and H3K27me3 occupancy in the CDKN2B promoter region. i ChIP assays were performed to detect EZH2 and H3K27me3 occupancy in the KLF2 promoter region. * P
    Figure Legend Snippet: SNHG1 participates in epigenetic repression of KLF2 and CDKN2B by interacting with PRC2. a GSEA showed a significant correlation between the SNHG1 and genes in PRC2 related pathway. b Scatter plot showing the expression relationship among SNHG1, EZH2, SUZ12 and EED in colorectal tumor tissues from TCGA database. The upper right squares show the Pearson correlation between each other. c RIPs experiments for EZH2, SUZ12 and EED were performed and the coprecipitated RNA was subjected to qRT-PCR for SNHG1. GAPDH was employed as a negative control. d RNA pull-down was used to examine the association of SNHG1 and EZH2. AR binding to HuR was used as a positive control. e PRC2 target genes expression was detected by qRT-PCR in SNHG1 siRNAs transfected HCT-116 cells. f CDKN2B and KLF2 expression was detected by qRT-PCR in SNHG1 vector transfected or SNHG1 vector and EZH2 siRNAs co-transfected CRC cells. g CDKN2B and KLF2 protein levels were detected by western blot in indicated conditions. h ChIP assays were performed to detect EZH2 and H3K27me3 occupancy in the CDKN2B promoter region. i ChIP assays were performed to detect EZH2 and H3K27me3 occupancy in the KLF2 promoter region. * P

    Techniques Used: Expressing, Quantitative RT-PCR, Negative Control, Binding Assay, Positive Control, Transfection, Plasmid Preparation, Western Blot, Chromatin Immunoprecipitation

    SNHG1 expression is up-regulated in colorectal cancer and is correlated with prognosis. a Hierarchical cluster heat map of differentially expressed lncRNAs in colorectal cancer and corresponding normal tissues generated from RNA sequencing data from the TCGA database. Red in the heat map denotes upregulation; blue denotes downregulation. The red arrow indicates SNHG1. b Expression of SNHG1 in the TCGA, GSE9348 and GSE8671 cohorts. c qRT-PCR analysis of SNHG1 expression in 80 pairs of colorectal cancer and corresponding normal tissues. d Kaplan-Meier survival analysis of CRC patients’ overall survival based on SNHG1 expression in our cohort ( n = 130, P
    Figure Legend Snippet: SNHG1 expression is up-regulated in colorectal cancer and is correlated with prognosis. a Hierarchical cluster heat map of differentially expressed lncRNAs in colorectal cancer and corresponding normal tissues generated from RNA sequencing data from the TCGA database. Red in the heat map denotes upregulation; blue denotes downregulation. The red arrow indicates SNHG1. b Expression of SNHG1 in the TCGA, GSE9348 and GSE8671 cohorts. c qRT-PCR analysis of SNHG1 expression in 80 pairs of colorectal cancer and corresponding normal tissues. d Kaplan-Meier survival analysis of CRC patients’ overall survival based on SNHG1 expression in our cohort ( n = 130, P

    Techniques Used: Expressing, Generated, RNA Sequencing Assay, Quantitative RT-PCR

    SP1 activates SNHG1 transcription in colorectal cancer cells. a Analysis of SP1 ChIP-seq, H3K4me3 ChIP-seq and DnaseI-seq data of HCT-116 cells in the SNHG1 locus. b SNHG1 expression was detected by qRT-PCR in HCT-116 and HCT-8 cells transfected with SP siRNAs or the SP1 vector. c The correlation between SP1 and SNHG1 expression analyzed in 30 paired colorectal cancer samples ( n = 30, r = 0.38, P = 0.03). d ChIP assays were performed to detect SP1 occupancy at the SNHG1 promoter region, α-Satellite and DHFR were employed as negative and positive control respectively for SP1 ChIP assays. e Dual luciferase reporter assays were used to determine the SP1 binding sites on the SNHG1 promoter region. The upper left corner of the picture was SP1 binding motif provided by the JASPAR CORE database. * P
    Figure Legend Snippet: SP1 activates SNHG1 transcription in colorectal cancer cells. a Analysis of SP1 ChIP-seq, H3K4me3 ChIP-seq and DnaseI-seq data of HCT-116 cells in the SNHG1 locus. b SNHG1 expression was detected by qRT-PCR in HCT-116 and HCT-8 cells transfected with SP siRNAs or the SP1 vector. c The correlation between SP1 and SNHG1 expression analyzed in 30 paired colorectal cancer samples ( n = 30, r = 0.38, P = 0.03). d ChIP assays were performed to detect SP1 occupancy at the SNHG1 promoter region, α-Satellite and DHFR were employed as negative and positive control respectively for SP1 ChIP assays. e Dual luciferase reporter assays were used to determine the SP1 binding sites on the SNHG1 promoter region. The upper left corner of the picture was SP1 binding motif provided by the JASPAR CORE database. * P

    Techniques Used: Chromatin Immunoprecipitation, Expressing, Quantitative RT-PCR, Transfection, Plasmid Preparation, Positive Control, Luciferase, Binding Assay

    SNHG1 regulates colorectal cancer cell proliferation and apoptosis. a Results of gene set enrichment analysis (GSEA) were plotted to visualize the correlation between the expression of SNHG1 and cell cycle and DNA repair gene signatures in TCGA cohort. b EdU assays were used to determine the cell proliferation ability of si-SNHG1 transfected cells. c Flow cytometric cell cycle distribution assays to detect the proportion of colorectal cancer cell cells in G1, S, and G2/M phases after transfection with SNHG1 siRNAs. d The cell cycle related proteins CyclinD1, CDK4, CDK6, and CyclinD2 were detected by western blot following SNHG1 silencing. e The effect of SNHG1 knockdown on cell apoptosis was analyzed by flow cytometric cell apoptosis assays. f Apoptosis related proteins Caspase-3, cleaved Caspase-3, PARP, cleaved PARP and Bax were detected by western blot after SNHG1 knockdown. Scale bar = 50 μm. ** P
    Figure Legend Snippet: SNHG1 regulates colorectal cancer cell proliferation and apoptosis. a Results of gene set enrichment analysis (GSEA) were plotted to visualize the correlation between the expression of SNHG1 and cell cycle and DNA repair gene signatures in TCGA cohort. b EdU assays were used to determine the cell proliferation ability of si-SNHG1 transfected cells. c Flow cytometric cell cycle distribution assays to detect the proportion of colorectal cancer cell cells in G1, S, and G2/M phases after transfection with SNHG1 siRNAs. d The cell cycle related proteins CyclinD1, CDK4, CDK6, and CyclinD2 were detected by western blot following SNHG1 silencing. e The effect of SNHG1 knockdown on cell apoptosis was analyzed by flow cytometric cell apoptosis assays. f Apoptosis related proteins Caspase-3, cleaved Caspase-3, PARP, cleaved PARP and Bax were detected by western blot after SNHG1 knockdown. Scale bar = 50 μm. ** P

    Techniques Used: Expressing, Transfection, Flow Cytometry, Western Blot

    SNHG1 affects colorectal cancer cells growth. a SNHG1 expression was detected by qRT-PCR in HCT-116 and HCT-8 cells transfected with two SNHG1 siRNAs. b HCT-116 and HCT-8 cells transfected with SNHG1 siRNAs were subjected to the CCK-8 assay after transfection. c HCT-116 and HCT-8 cells transfected with SNHG1 siRNAs were seeded onto 6-well plates. The number of colonies was counted on the 14th day after seeding. d Representative images of mice bearing tumors from empty vector, sh-SNHG1#1 vector and SNHG1 vector groups, and the tumor volume growth curves after injections in different groups. e SNHG1 expression was detected in tumors from different groups of mice using qRT-PCR. f Representative images of hematoxylin and eosin (HE) staining and Ki67 immunostaining of tumor samples from different groups. Scale bar = 50 μm. * P
    Figure Legend Snippet: SNHG1 affects colorectal cancer cells growth. a SNHG1 expression was detected by qRT-PCR in HCT-116 and HCT-8 cells transfected with two SNHG1 siRNAs. b HCT-116 and HCT-8 cells transfected with SNHG1 siRNAs were subjected to the CCK-8 assay after transfection. c HCT-116 and HCT-8 cells transfected with SNHG1 siRNAs were seeded onto 6-well plates. The number of colonies was counted on the 14th day after seeding. d Representative images of mice bearing tumors from empty vector, sh-SNHG1#1 vector and SNHG1 vector groups, and the tumor volume growth curves after injections in different groups. e SNHG1 expression was detected in tumors from different groups of mice using qRT-PCR. f Representative images of hematoxylin and eosin (HE) staining and Ki67 immunostaining of tumor samples from different groups. Scale bar = 50 μm. * P

    Techniques Used: Expressing, Quantitative RT-PCR, Transfection, CCK-8 Assay, Mouse Assay, Plasmid Preparation, Staining, Immunostaining

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    ATCC treatment human colorectal cancer cell lines
    <t>Anti-colorectal</t> cancer effects of curcumin and/or irinotecan are dependent on ROS ( A , B ) The effects of NAC on cell growth inhibition induced by curcumin and/or irinotecan. After pretreatment with 5 mM NAC for 2 h, LoVo cells (A) or HT-29 cells (B) were treated with curcumin and/or irinotecan for 24 h, then cell viability was assessed by CCK-8 assay. ( C , D ) The effects of NAC on apoptosis induced by curcumin and/or irinotecan. After cells were treated as described above, cell apoptosis was measured by Annexin V-FITC/PI staining. Values are means ± SEM. *
    Treatment Human Colorectal Cancer Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/treatment human colorectal cancer cell lines/product/ATCC
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    treatment human colorectal cancer cell lines - by Bioz Stars, 2022-09
    90/100 stars
      Buy from Supplier

    88
    ATCC human crc cell lines
    miR-150-5p inhibited <t>CRC</t> progression by targeting VEGFA. ( A ) VEGFA protein expression was determined in <t>HCT116</t> and HCT8 cells transfected with agomiR-150-5p with VEGFA expression plasmid or empty vector using western blot; GAPDH was used as the internal control. ( B - D ) Cell proliferation ( B , C ), migration ( D )and invasion ( E ) were evaluated in HCT116 and HCT8 cells transfected with agomiR-150-5p with VEGFA expression plasmid or empty vector. ( F ) HUVECs were cultured in TCM derived from HCT116 and HCT8 cells transfected with agomiR-150-5p plus VEGFA expression plasmid or empty vector. Data are shown as the mean±SD. * p
    Human Crc Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human crc cell lines/product/ATCC
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human crc cell lines - by Bioz Stars, 2022-09
    88/100 stars
      Buy from Supplier

    90
    ATCC human colorectal cancer cell lines
    Endogenous expression of Pdcd4, CD24, Src, miR-21 and miR-34a in resected <t>colorectal</t> tissues. ( a ) Western blot analysis was performed for Pdcd4, CD24 and Src in colorectal tumors (Tumor) and corresponding normal tissues (Normal) taken from a series of 26 patients. β-Actin served as internal control. Relative mean protein amounts (Fold change comparative to normal tissue expression) of Pdcd4, CD24 and Src obtained by densitometry analysis are represented as bar graphs. Specific Pdcd4, CD24 or Src band intersities were normalized with β-actin. Pdcd4 was downregulated, CD24 and Src were upregulated significantly in the tumor tissues (p = 0.003, p = 0.05 and p = 0.001, respectively) ( b ) Real-time PCR results of miR-21 and miR-34a in the same colorectal tumor (Tumor) and normal tissue (Normal) samples. Mean relative expression (fold change compared to expression in normal tissue) of miR-21 and miR-34a is represented as bar graphs. miR-21 was upregulated and miR-34a was downregulated significantly in the tumor tissues. (p = 0.002, p = 0.05, respectively) ( c ) Lysates from 7 representative normal tissue (N) and colorectal tumor (T) samples were subjected to Western blotting and probed for the expression of Pdcd4, CD24 and Src and represented. β-Actin served as a loading control ( d ) Schematic representation of the functional network between CD24, Src, AP-1, miR-21, Pdcd4 and miR-34a.
    Human Colorectal Cancer Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human colorectal cancer cell lines/product/ATCC
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human colorectal cancer cell lines - by Bioz Stars, 2022-09
    90/100 stars
      Buy from Supplier

    99
    ATCC 293ft human embryonic kidney cell line
    PRL-1 is not a direct target of miR-339-3p in CRC cells. (A) The expression of PRL-1 mRNAs were analysis by reverse transcription-quantitative polymerase chain reaction. (B) The protein expression levels of PRL-1 were detected using western blot analysis. (C) Analysis of luciferase activity. <t>293FT</t> cells and HCT116 cells were co-transfected with psiCHECK™-2 luciferase reporter plasmid containing either wt or mut PRL-1 3′-UTR and either the miR-339-3p mimics or non-specific miR mimic control (NC). Luciferase activity was assayed 48 h after transfection. Renilla luciferase activity of each sample was normalized by Firefly luciferase activity. Data are presented as mean ± SD from 3 independent experiments. CRC, colorectal cancer; wt, wild-type; mut, mutant.
    293ft Human Embryonic Kidney Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/293ft human embryonic kidney cell line/product/ATCC
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    293ft human embryonic kidney cell line - by Bioz Stars, 2022-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Anti-colorectal cancer effects of curcumin and/or irinotecan are dependent on ROS ( A , B ) The effects of NAC on cell growth inhibition induced by curcumin and/or irinotecan. After pretreatment with 5 mM NAC for 2 h, LoVo cells (A) or HT-29 cells (B) were treated with curcumin and/or irinotecan for 24 h, then cell viability was assessed by CCK-8 assay. ( C , D ) The effects of NAC on apoptosis induced by curcumin and/or irinotecan. After cells were treated as described above, cell apoptosis was measured by Annexin V-FITC/PI staining. Values are means ± SEM. *

    Journal: Oncotarget

    Article Title: Curcumin enhances the effects of irinotecan on colorectal cancer cells through the generation of reactive oxygen species and activation of the endoplasmic reticulum stress pathway

    doi: 10.18632/oncotarget.16828

    Figure Lengend Snippet: Anti-colorectal cancer effects of curcumin and/or irinotecan are dependent on ROS ( A , B ) The effects of NAC on cell growth inhibition induced by curcumin and/or irinotecan. After pretreatment with 5 mM NAC for 2 h, LoVo cells (A) or HT-29 cells (B) were treated with curcumin and/or irinotecan for 24 h, then cell viability was assessed by CCK-8 assay. ( C , D ) The effects of NAC on apoptosis induced by curcumin and/or irinotecan. After cells were treated as described above, cell apoptosis was measured by Annexin V-FITC/PI staining. Values are means ± SEM. *

    Article Snippet: Cell culture and treatment Human colorectal cancer cell lines, LoVo and HT-29, were obtained from the American Type Culture Collection (Manassas, VA, USA).

    Techniques: Inhibition, CCK-8 Assay, Staining

    ER Stress is mediates the anti-colorectal cancer effects of curcumin alone or combined with irinotecan ( A , B ) The effects of an ER stress inhibitor on cell growth inhibition induced by curcumin alone or with irinotecan. After pretreatment with 0.1 μM mithramycin (MTM) for 30 min, LoVo cells (A) or HT-29 cells (B) were treated with curcumin alone or with irinotecan for 24 h, then cell viability was assessed by CCK-8 assay. ( C , D ) The effects of an ER stress inhibitor on apoptosis induced by curcumin alone or with irinotecan. After cells were treated as described above, cell apoptosis was measured by Annexin V-FITC/PI staining. Values are means ± SEM. *

    Journal: Oncotarget

    Article Title: Curcumin enhances the effects of irinotecan on colorectal cancer cells through the generation of reactive oxygen species and activation of the endoplasmic reticulum stress pathway

    doi: 10.18632/oncotarget.16828

    Figure Lengend Snippet: ER Stress is mediates the anti-colorectal cancer effects of curcumin alone or combined with irinotecan ( A , B ) The effects of an ER stress inhibitor on cell growth inhibition induced by curcumin alone or with irinotecan. After pretreatment with 0.1 μM mithramycin (MTM) for 30 min, LoVo cells (A) or HT-29 cells (B) were treated with curcumin alone or with irinotecan for 24 h, then cell viability was assessed by CCK-8 assay. ( C , D ) The effects of an ER stress inhibitor on apoptosis induced by curcumin alone or with irinotecan. After cells were treated as described above, cell apoptosis was measured by Annexin V-FITC/PI staining. Values are means ± SEM. *

    Article Snippet: Cell culture and treatment Human colorectal cancer cell lines, LoVo and HT-29, were obtained from the American Type Culture Collection (Manassas, VA, USA).

    Techniques: Inhibition, CCK-8 Assay, Staining

    miR-150-5p inhibited CRC progression by targeting VEGFA. ( A ) VEGFA protein expression was determined in HCT116 and HCT8 cells transfected with agomiR-150-5p with VEGFA expression plasmid or empty vector using western blot; GAPDH was used as the internal control. ( B - D ) Cell proliferation ( B , C ), migration ( D )and invasion ( E ) were evaluated in HCT116 and HCT8 cells transfected with agomiR-150-5p with VEGFA expression plasmid or empty vector. ( F ) HUVECs were cultured in TCM derived from HCT116 and HCT8 cells transfected with agomiR-150-5p plus VEGFA expression plasmid or empty vector. Data are shown as the mean±SD. * p

    Journal: Aging (Albany NY)

    Article Title: miR-150-5p suppresses tumor progression by targeting VEGFA in colorectal cancer

    doi: 10.18632/aging.101656

    Figure Lengend Snippet: miR-150-5p inhibited CRC progression by targeting VEGFA. ( A ) VEGFA protein expression was determined in HCT116 and HCT8 cells transfected with agomiR-150-5p with VEGFA expression plasmid or empty vector using western blot; GAPDH was used as the internal control. ( B - D ) Cell proliferation ( B , C ), migration ( D )and invasion ( E ) were evaluated in HCT116 and HCT8 cells transfected with agomiR-150-5p with VEGFA expression plasmid or empty vector. ( F ) HUVECs were cultured in TCM derived from HCT116 and HCT8 cells transfected with agomiR-150-5p plus VEGFA expression plasmid or empty vector. Data are shown as the mean±SD. * p

    Article Snippet: The human CRC cell lines including HCT116, HCT8, HT29, SW620, SW480 and DLD-1 and normal colonic epithelial cells (FHC) were obtained from ATCC.

    Techniques: Expressing, Transfection, Plasmid Preparation, Western Blot, Migration, Cell Culture, Derivative Assay

    A. VEGFA was a direct target of miR-150-5p in CRC. ( A ) The direct target genes of miR-150-5p were predicted using the PicTarSites, miRandaSites and Tarbase databases. ( B ) Wild-type and mutant VEGFA-3’UTR sequences were cloned into luciferase reporter. Luciferase activity was determined in HCT116 and 293T cells cotransfected with agomiR-150-5p or agomiR-NC and pmirGLO-VEGFA-3’UTR-WT or pmirGLO-VEGFA-3’UTR-Mut. Luciferase activities were normalized to that of renilla luciferase. C, D. qRT-PCR ( C ) and western blot ( D ) analyses showed that both VEGFA mRNA and protein expression levels were dramatically suppressed by agomiR-150-5p in HCT116 and HCT8 cells, GAPDH was used as the internal control. ** p

    Journal: Aging (Albany NY)

    Article Title: miR-150-5p suppresses tumor progression by targeting VEGFA in colorectal cancer

    doi: 10.18632/aging.101656

    Figure Lengend Snippet: A. VEGFA was a direct target of miR-150-5p in CRC. ( A ) The direct target genes of miR-150-5p were predicted using the PicTarSites, miRandaSites and Tarbase databases. ( B ) Wild-type and mutant VEGFA-3’UTR sequences were cloned into luciferase reporter. Luciferase activity was determined in HCT116 and 293T cells cotransfected with agomiR-150-5p or agomiR-NC and pmirGLO-VEGFA-3’UTR-WT or pmirGLO-VEGFA-3’UTR-Mut. Luciferase activities were normalized to that of renilla luciferase. C, D. qRT-PCR ( C ) and western blot ( D ) analyses showed that both VEGFA mRNA and protein expression levels were dramatically suppressed by agomiR-150-5p in HCT116 and HCT8 cells, GAPDH was used as the internal control. ** p

    Article Snippet: The human CRC cell lines including HCT116, HCT8, HT29, SW620, SW480 and DLD-1 and normal colonic epithelial cells (FHC) were obtained from ATCC.

    Techniques: Mutagenesis, Clone Assay, Luciferase, Activity Assay, Quantitative RT-PCR, Western Blot, Expressing

    VEGFA knockdown significantly inhibited CRC progression. ( A ) VEGFA expression was downregulated in HCT116 and HCT8 cells transfected with siVEGFA-1 or siVEGFA-2. ( B ) VEGFA knockdown inhibited CRC cell proliferation ( B ), migration ( C ), invasion ( D ) and HUVECs tube formation ( E ). Data are shown as the mean±SD of three independent experiments. * p

    Journal: Aging (Albany NY)

    Article Title: miR-150-5p suppresses tumor progression by targeting VEGFA in colorectal cancer

    doi: 10.18632/aging.101656

    Figure Lengend Snippet: VEGFA knockdown significantly inhibited CRC progression. ( A ) VEGFA expression was downregulated in HCT116 and HCT8 cells transfected with siVEGFA-1 or siVEGFA-2. ( B ) VEGFA knockdown inhibited CRC cell proliferation ( B ), migration ( C ), invasion ( D ) and HUVECs tube formation ( E ). Data are shown as the mean±SD of three independent experiments. * p

    Article Snippet: The human CRC cell lines including HCT116, HCT8, HT29, SW620, SW480 and DLD-1 and normal colonic epithelial cells (FHC) were obtained from ATCC.

    Techniques: Expressing, Transfection, Migration

    Endogenous expression of Pdcd4, CD24, Src, miR-21 and miR-34a in resected colorectal tissues. ( a ) Western blot analysis was performed for Pdcd4, CD24 and Src in colorectal tumors (Tumor) and corresponding normal tissues (Normal) taken from a series of 26 patients. β-Actin served as internal control. Relative mean protein amounts (Fold change comparative to normal tissue expression) of Pdcd4, CD24 and Src obtained by densitometry analysis are represented as bar graphs. Specific Pdcd4, CD24 or Src band intersities were normalized with β-actin. Pdcd4 was downregulated, CD24 and Src were upregulated significantly in the tumor tissues (p = 0.003, p = 0.05 and p = 0.001, respectively) ( b ) Real-time PCR results of miR-21 and miR-34a in the same colorectal tumor (Tumor) and normal tissue (Normal) samples. Mean relative expression (fold change compared to expression in normal tissue) of miR-21 and miR-34a is represented as bar graphs. miR-21 was upregulated and miR-34a was downregulated significantly in the tumor tissues. (p = 0.002, p = 0.05, respectively) ( c ) Lysates from 7 representative normal tissue (N) and colorectal tumor (T) samples were subjected to Western blotting and probed for the expression of Pdcd4, CD24 and Src and represented. β-Actin served as a loading control ( d ) Schematic representation of the functional network between CD24, Src, AP-1, miR-21, Pdcd4 and miR-34a.

    Journal: PLoS ONE

    Article Title: CD24 Induces Expression of the Oncomir miR-21 via Src, and CD24 and Src Are Both Post-Transcriptionally Downregulated by the Tumor Suppressor miR-34a

    doi: 10.1371/journal.pone.0059563

    Figure Lengend Snippet: Endogenous expression of Pdcd4, CD24, Src, miR-21 and miR-34a in resected colorectal tissues. ( a ) Western blot analysis was performed for Pdcd4, CD24 and Src in colorectal tumors (Tumor) and corresponding normal tissues (Normal) taken from a series of 26 patients. β-Actin served as internal control. Relative mean protein amounts (Fold change comparative to normal tissue expression) of Pdcd4, CD24 and Src obtained by densitometry analysis are represented as bar graphs. Specific Pdcd4, CD24 or Src band intersities were normalized with β-actin. Pdcd4 was downregulated, CD24 and Src were upregulated significantly in the tumor tissues (p = 0.003, p = 0.05 and p = 0.001, respectively) ( b ) Real-time PCR results of miR-21 and miR-34a in the same colorectal tumor (Tumor) and normal tissue (Normal) samples. Mean relative expression (fold change compared to expression in normal tissue) of miR-21 and miR-34a is represented as bar graphs. miR-21 was upregulated and miR-34a was downregulated significantly in the tumor tissues. (p = 0.002, p = 0.05, respectively) ( c ) Lysates from 7 representative normal tissue (N) and colorectal tumor (T) samples were subjected to Western blotting and probed for the expression of Pdcd4, CD24 and Src and represented. β-Actin served as a loading control ( d ) Schematic representation of the functional network between CD24, Src, AP-1, miR-21, Pdcd4 and miR-34a.

    Article Snippet: Cell Culture and Antibodies The human colorectal cancer cell lines (HT-29, HCT-116, Rko, SW480, Colo206f and WiDr) and the human breast cancer cell line MDA-MB-231 were purchased from American Type Culture Collection (ATCC, Manassas, USA), and grown according to the recommended conditions.

    Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Functional Assay

    PRL-1 is not a direct target of miR-339-3p in CRC cells. (A) The expression of PRL-1 mRNAs were analysis by reverse transcription-quantitative polymerase chain reaction. (B) The protein expression levels of PRL-1 were detected using western blot analysis. (C) Analysis of luciferase activity. 293FT cells and HCT116 cells were co-transfected with psiCHECK™-2 luciferase reporter plasmid containing either wt or mut PRL-1 3′-UTR and either the miR-339-3p mimics or non-specific miR mimic control (NC). Luciferase activity was assayed 48 h after transfection. Renilla luciferase activity of each sample was normalized by Firefly luciferase activity. Data are presented as mean ± SD from 3 independent experiments. CRC, colorectal cancer; wt, wild-type; mut, mutant.

    Journal: Oncology Letters

    Article Title: miR-339-3p inhibits proliferation and metastasis of colorectal cancer

    doi: 10.3892/ol.2015.3661

    Figure Lengend Snippet: PRL-1 is not a direct target of miR-339-3p in CRC cells. (A) The expression of PRL-1 mRNAs were analysis by reverse transcription-quantitative polymerase chain reaction. (B) The protein expression levels of PRL-1 were detected using western blot analysis. (C) Analysis of luciferase activity. 293FT cells and HCT116 cells were co-transfected with psiCHECK™-2 luciferase reporter plasmid containing either wt or mut PRL-1 3′-UTR and either the miR-339-3p mimics or non-specific miR mimic control (NC). Luciferase activity was assayed 48 h after transfection. Renilla luciferase activity of each sample was normalized by Firefly luciferase activity. Data are presented as mean ± SD from 3 independent experiments. CRC, colorectal cancer; wt, wild-type; mut, mutant.

    Article Snippet: Cell culture and miRNA transfection The 293FT human embryonic kidney cell line and 6 human colorectal cancer cell lines (HCT116, HT29, LS174T, SW480, SW620 and LOVO) with different metastatic abilities were purchased from The American Type Culture Collection (ATCC, Manassas, VA, USA).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Mutagenesis