colorectal cancer cell lines  (ATCC)


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    Structured Review

    ATCC colorectal cancer cell lines
    Expression of PRDX2 is elevated in primary <t>colorectal</t> tumors compared with human colorectal tumor-adjacent tissues ( A ) Expression of PRDX2 protein in each of the primary colorectal tumors and colorectal adjacent noncancerous tissues paired from the same patient by Western blotting, GAPDH was used as loading control. ( B ) Expression of PRDX2 mRNA in each of the primary colorectal tumors (T) and colorectal adjacent noncancerous tissues (ANT) paired from the same patient by quantitative real-time reverse transcription-PCR (qRT-PCR). ( C ) PRDX2 expression level was up-regulated in the primary colorectal tumor compared with the paired colorectal adjacent noncancerous tissues from the same patient, as examined by immunohistochemistry. Data represent the mean ± SD of three experiments. (* p
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    Images

    1) Product Images from "Peroxiredoxin 2 is associated with colorectal cancer progression and poor survival of patients"

    Article Title: Peroxiredoxin 2 is associated with colorectal cancer progression and poor survival of patients

    Journal: Oncotarget

    doi: 10.18632/oncotarget.14801

    Expression of PRDX2 is elevated in primary colorectal tumors compared with human colorectal tumor-adjacent tissues ( A ) Expression of PRDX2 protein in each of the primary colorectal tumors and colorectal adjacent noncancerous tissues paired from the same patient by Western blotting, GAPDH was used as loading control. ( B ) Expression of PRDX2 mRNA in each of the primary colorectal tumors (T) and colorectal adjacent noncancerous tissues (ANT) paired from the same patient by quantitative real-time reverse transcription-PCR (qRT-PCR). ( C ) PRDX2 expression level was up-regulated in the primary colorectal tumor compared with the paired colorectal adjacent noncancerous tissues from the same patient, as examined by immunohistochemistry. Data represent the mean ± SD of three experiments. (* p
    Figure Legend Snippet: Expression of PRDX2 is elevated in primary colorectal tumors compared with human colorectal tumor-adjacent tissues ( A ) Expression of PRDX2 protein in each of the primary colorectal tumors and colorectal adjacent noncancerous tissues paired from the same patient by Western blotting, GAPDH was used as loading control. ( B ) Expression of PRDX2 mRNA in each of the primary colorectal tumors (T) and colorectal adjacent noncancerous tissues (ANT) paired from the same patient by quantitative real-time reverse transcription-PCR (qRT-PCR). ( C ) PRDX2 expression level was up-regulated in the primary colorectal tumor compared with the paired colorectal adjacent noncancerous tissues from the same patient, as examined by immunohistochemistry. Data represent the mean ± SD of three experiments. (* p

    Techniques Used: Expressing, Western Blot, Polymerase Chain Reaction, Quantitative RT-PCR, Immunohistochemistry

    Expression analysis of PRDX2 protein and mRNA in HCEC and colorectal cancer cell lines by quantitative real-time reverse transcription-PCR (qRT-PCR) and Western blotting ( A ) Expression of PRDX2 protein in HCEC and cultured colorectal cancer cell lines SW480, SW620, HT29, HCT116, and SW837. ( B ) Expression of PRDX2 mRNA in HCEC and cultured colorectal cancer cell lines. Data represent the mean ± SD of three experiments. (* p
    Figure Legend Snippet: Expression analysis of PRDX2 protein and mRNA in HCEC and colorectal cancer cell lines by quantitative real-time reverse transcription-PCR (qRT-PCR) and Western blotting ( A ) Expression of PRDX2 protein in HCEC and cultured colorectal cancer cell lines SW480, SW620, HT29, HCT116, and SW837. ( B ) Expression of PRDX2 mRNA in HCEC and cultured colorectal cancer cell lines. Data represent the mean ± SD of three experiments. (* p

    Techniques Used: Expressing, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Cell Culture

    PRDX2 is frequently up-regulated in CRC ( A ) Representative immunohistochemical expression patterns of PRDX2 in 226 paired human primary colorectal cancer tissues and corresponding adjacent normal mucosa specimens are shown. ( B ) Percentage of cases with different staining intensity of PRDX2 in the tumor or adjacent normal tissues in the study cohort.
    Figure Legend Snippet: PRDX2 is frequently up-regulated in CRC ( A ) Representative immunohistochemical expression patterns of PRDX2 in 226 paired human primary colorectal cancer tissues and corresponding adjacent normal mucosa specimens are shown. ( B ) Percentage of cases with different staining intensity of PRDX2 in the tumor or adjacent normal tissues in the study cohort.

    Techniques Used: Immunohistochemistry, Expressing, Staining

    2) Product Images from "SPRY4 suppresses proliferation and induces apoptosis of colorectal cancer cells by repressing oncogene EZH2"

    Article Title: SPRY4 suppresses proliferation and induces apoptosis of colorectal cancer cells by repressing oncogene EZH2

    Journal: Aging (Albany NY)

    doi: 10.18632/aging.202859

    Differential expression of SPRY4 and EZH2 between colorectal cancer cells and normal human colon cell. From four human CRC cell lines, SW620, SW480, LOVO and HCT116, and a normal human colon mucosal epithelial cell line NCM460, we detected the mRNA level and protein expression of SPRY4 using western blotting and RT-qPCR, respectively ( A – C ). The mRNA level and protein expression of EZH2 were detected using western blotting and RT-qPCR, respectively ( D – F ). **, ***p
    Figure Legend Snippet: Differential expression of SPRY4 and EZH2 between colorectal cancer cells and normal human colon cell. From four human CRC cell lines, SW620, SW480, LOVO and HCT116, and a normal human colon mucosal epithelial cell line NCM460, we detected the mRNA level and protein expression of SPRY4 using western blotting and RT-qPCR, respectively ( A – C ). The mRNA level and protein expression of EZH2 were detected using western blotting and RT-qPCR, respectively ( D – F ). **, ***p

    Techniques Used: Expressing, Western Blot, Quantitative RT-PCR

    3) Product Images from "A functional screen with metformin identifies microRNAs that regulate metabolism in colorectal cancer cells"

    Article Title: A functional screen with metformin identifies microRNAs that regulate metabolism in colorectal cancer cells

    Journal: Scientific Reports

    doi: 10.1038/s41598-022-06587-9

    Changes in the reserved respiratory capacity ( a ), coupling efficiency ( b ), basal respiration ( c ), maximal respiratory capacity ( d ) and ATP synthesis ( e ) of HCT116 cells associated with miRNA mimic transfections in combination with metformin treatment. CRC cells were reverse transfected with selected miRNA mimics and treated with 2.5 mM metformin treatment or control medium. Oxygen consumption rate (OCR) of the cells was determined over a 30-min period using Seahorse XFe-96 instrument. The OCR values were normalized to viability of the corresponding cells acquired by crystal violet assay. Results are expressed as mean ± SD of 3 culture replicates and the statistical significance is indicated with asterisks (ns P > 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 and ****P ≤ 0.0001).
    Figure Legend Snippet: Changes in the reserved respiratory capacity ( a ), coupling efficiency ( b ), basal respiration ( c ), maximal respiratory capacity ( d ) and ATP synthesis ( e ) of HCT116 cells associated with miRNA mimic transfections in combination with metformin treatment. CRC cells were reverse transfected with selected miRNA mimics and treated with 2.5 mM metformin treatment or control medium. Oxygen consumption rate (OCR) of the cells was determined over a 30-min period using Seahorse XFe-96 instrument. The OCR values were normalized to viability of the corresponding cells acquired by crystal violet assay. Results are expressed as mean ± SD of 3 culture replicates and the statistical significance is indicated with asterisks (ns P > 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 and ****P ≤ 0.0001).

    Techniques Used: Transfection, Crystal Violet Assay

    Viability of HCT116 cells transfected with miRNA mimics that show a strong synergetic effect with 2.5 mM metformin treatment. Normalized cell counts of HCT116 cells transfected with miRNA mimics, treated with 2.5 mM metformin and compared with 0 mM metformin treatment. Results are expressed as mean ± SD of 3 replicates and the statistical significance is indicated with asterisks (ns P > 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 and ****P ≤ 0.0001).
    Figure Legend Snippet: Viability of HCT116 cells transfected with miRNA mimics that show a strong synergetic effect with 2.5 mM metformin treatment. Normalized cell counts of HCT116 cells transfected with miRNA mimics, treated with 2.5 mM metformin and compared with 0 mM metformin treatment. Results are expressed as mean ± SD of 3 replicates and the statistical significance is indicated with asterisks (ns P > 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 and ****P ≤ 0.0001).

    Techniques Used: Transfection

    Changes in glycolysis ( a ), maximum glycolytic capacity ( b ) and glycolytic reserve ( c ) associated with miRNA mimic transfections in combination with metformin treatment of HCT116 cells. CRC cells were reverse transfected with selected miRNA mimics and treated with 2.5 mM metformin or control medium. Extracellular acidification rate (ECAR) of the cells was determined over a 30-min period using a Seahorse XFe-96 Instrument. The ECAR values were normalized to viability of the cells per group determined by crystal violet assay. Results are expressed as mean ± SD of 3 culture replicates and the statistical significance is indicated with asterisks (ns P > 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001.
    Figure Legend Snippet: Changes in glycolysis ( a ), maximum glycolytic capacity ( b ) and glycolytic reserve ( c ) associated with miRNA mimic transfections in combination with metformin treatment of HCT116 cells. CRC cells were reverse transfected with selected miRNA mimics and treated with 2.5 mM metformin or control medium. Extracellular acidification rate (ECAR) of the cells was determined over a 30-min period using a Seahorse XFe-96 Instrument. The ECAR values were normalized to viability of the cells per group determined by crystal violet assay. Results are expressed as mean ± SD of 3 culture replicates and the statistical significance is indicated with asterisks (ns P > 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001.

    Techniques Used: Transfection, Crystal Violet Assay

    Integrated energy maps of HCT116 cells associated with miRNA mimic transfections in combination with metformin treatment. CRC cells were reverse transfected with selected miRNA mimics and treated with 2.5 mM metformin treatment or control medium. The baseline oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) calculated were combined to generate the basal phenogram. The OCR and ECAR values were normalized to viability of the corresponding cells acquired by crystal violet assay. Results are expressed as mean ± SD of 3 technical replicates.
    Figure Legend Snippet: Integrated energy maps of HCT116 cells associated with miRNA mimic transfections in combination with metformin treatment. CRC cells were reverse transfected with selected miRNA mimics and treated with 2.5 mM metformin treatment or control medium. The baseline oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) calculated were combined to generate the basal phenogram. The OCR and ECAR values were normalized to viability of the corresponding cells acquired by crystal violet assay. Results are expressed as mean ± SD of 3 technical replicates.

    Techniques Used: Transfection, Crystal Violet Assay

    Schematic of high throughput screen of miRNA mimics, summary of data for CRC cell response to metformin treatment and scatter plot representation of the metformin sensitization screen with a miRNA mimic library. ( a ) Workflow of functional miRNA mimic screen is demonstrated. HCT116 cells were reverse transfected with 25 nM miRNA mimics and following 24 h incubation, cells were treated with 2.5 mM metformin or control medium for 72 h. Endpoint high-content image analysis was performed for cells stained with Hoechst 33342 and Calcein-AM. ( b ) Summary of viability data for the functional high throughput screen of miRNA mimics in combination with metformin is shown. Negative control for cell death is represented by the viability of cells transfected with negative control (OTP) and positive control is represented by the cells transfected with siRNA targeting PLK1. Mock control also shows the control for transfection ( c ) Percent viability of HCT116 cells reverse transfected with miRNA mimics and treated with 2.5 mM (y- axis) or 0 mM metformin (x-axis) are shown. The viability values were normalized against siOTP values and miRNAs selected for a secondary screen (blue triangles).
    Figure Legend Snippet: Schematic of high throughput screen of miRNA mimics, summary of data for CRC cell response to metformin treatment and scatter plot representation of the metformin sensitization screen with a miRNA mimic library. ( a ) Workflow of functional miRNA mimic screen is demonstrated. HCT116 cells were reverse transfected with 25 nM miRNA mimics and following 24 h incubation, cells were treated with 2.5 mM metformin or control medium for 72 h. Endpoint high-content image analysis was performed for cells stained with Hoechst 33342 and Calcein-AM. ( b ) Summary of viability data for the functional high throughput screen of miRNA mimics in combination with metformin is shown. Negative control for cell death is represented by the viability of cells transfected with negative control (OTP) and positive control is represented by the cells transfected with siRNA targeting PLK1. Mock control also shows the control for transfection ( c ) Percent viability of HCT116 cells reverse transfected with miRNA mimics and treated with 2.5 mM (y- axis) or 0 mM metformin (x-axis) are shown. The viability values were normalized against siOTP values and miRNAs selected for a secondary screen (blue triangles).

    Techniques Used: High Throughput Screening Assay, Functional Assay, Transfection, Incubation, Staining, Negative Control, Positive Control

    Cell cycle distribution of HCT116 cells after transfection with sensitizing miRNAs and treatment with metformin or control medium for 96 h ( a ) and the miRNA-based sub-network of putative target genes associated with cell cycle ( b ). ( a ) Histograms showing the percentage of HCT116 cells transfected with miR-18b-5p, miR-145-3p, miR-376b-5p, miR-676-3p and miR-718 mimics and treated with 2.5 mM metformin for 72 h in G0/G1, S, and G2/M phases, compared with cells in control medium. A “–” miRNA mimic status indicates transfection with NC negative control mimics. Results are expressed as mean ± SD of 3 replicates and the statistical significance is indicated with asterisks (ns P > 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001). ( b ) This is an organic layout of the integrated network associated putative genes organised in cell cycle associated ontologies acquired by Targetscan predition algorithm and NetworkAnalyst-Panther/Slim GO analysis. The miRNA-gene interactions are shown as red lines while gene–gene interactions are green. The blue and red nodes represent predicted taget genes and miRNAs, respectively.
    Figure Legend Snippet: Cell cycle distribution of HCT116 cells after transfection with sensitizing miRNAs and treatment with metformin or control medium for 96 h ( a ) and the miRNA-based sub-network of putative target genes associated with cell cycle ( b ). ( a ) Histograms showing the percentage of HCT116 cells transfected with miR-18b-5p, miR-145-3p, miR-376b-5p, miR-676-3p and miR-718 mimics and treated with 2.5 mM metformin for 72 h in G0/G1, S, and G2/M phases, compared with cells in control medium. A “–” miRNA mimic status indicates transfection with NC negative control mimics. Results are expressed as mean ± SD of 3 replicates and the statistical significance is indicated with asterisks (ns P > 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001). ( b ) This is an organic layout of the integrated network associated putative genes organised in cell cycle associated ontologies acquired by Targetscan predition algorithm and NetworkAnalyst-Panther/Slim GO analysis. The miRNA-gene interactions are shown as red lines while gene–gene interactions are green. The blue and red nodes represent predicted taget genes and miRNAs, respectively.

    Techniques Used: Transfection, Negative Control

    4) Product Images from "hSETD1A Regulates Wnt Target Genes and Controls Tumor Growth of Colorectal Cancer Cells"

    Article Title: hSETD1A Regulates Wnt Target Genes and Controls Tumor Growth of Colorectal Cancer Cells

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-13-1400

    A working model illustrating the cooperative role of hSETD1A and β-catenin in regulation of Wnt target genes in colorectal cancer. In normal colon cells, β-catenin levels remain low due to its phosphorylation and subsequent degradation. In colorectal cancer, both hSETD1A and β-catenin levels are upregulated. Localization of β-catenin to the nucleus enables its interactions with hSETD1A HMT. These interactions facilitate promoter-H3K4me3, recruitment of TAF3 (and transcription machinery), activation of Wnt/β-catenin target genes, and cellular growth.
    Figure Legend Snippet: A working model illustrating the cooperative role of hSETD1A and β-catenin in regulation of Wnt target genes in colorectal cancer. In normal colon cells, β-catenin levels remain low due to its phosphorylation and subsequent degradation. In colorectal cancer, both hSETD1A and β-catenin levels are upregulated. Localization of β-catenin to the nucleus enables its interactions with hSETD1A HMT. These interactions facilitate promoter-H3K4me3, recruitment of TAF3 (and transcription machinery), activation of Wnt/β-catenin target genes, and cellular growth.

    Techniques Used: HMT Assay, Activation Assay

    hSETD1A interacts with β-catenin and regulates H3K4me3 at promoters of Wnt target genes. A, the levels of hSETD1A and selected Wnt target genes were analyzed by Western blot (WB) analysis. β-Catenin is unaffected by the knockdown of hSETD1A in HCT116 cells. B and C, coimmunoprecipitation assays revealed that hSETD1A interacts with β-catenin in HCT116 (B) and SW48 (C) cells. D, immunofluorescent staining of colorectal tumor and paired normal mucosa from patient #2 revealed that β-catenin is highly expressed in the intestinal crypt. E, the hSETD1A complex colocalizes with the β-catenin complex at the known β-catenin responsive element at MYC gene in HCT116 cells. ChIP analysis was performed using antibodies against hSETD1A complex (hSETD1A, Ash2L, WDR5, and RbBP5) and β-catenin complex (β-catenin and TCF4) to determine their binding to the known β-catenin responsive element at MYC enhancer and a negative control region located 70 Kb downstream the TAL1 promoter in HCT116. F and G, H3K4me3 (F), and β-catenin (G) ChIP assays were performed in the Scramble (Scr) control and hSETD1A knockdown HCT116 cells. H3K4me3 enrichment was decreased at the TSSs of the selected Wnt target genes upon hSETD1A knockdown, whereas β-catenin binding remained unchanged. The VEGFA 3′UTR was shown as a negative control region. Data, mean ± SD from three independent replicates. *, P
    Figure Legend Snippet: hSETD1A interacts with β-catenin and regulates H3K4me3 at promoters of Wnt target genes. A, the levels of hSETD1A and selected Wnt target genes were analyzed by Western blot (WB) analysis. β-Catenin is unaffected by the knockdown of hSETD1A in HCT116 cells. B and C, coimmunoprecipitation assays revealed that hSETD1A interacts with β-catenin in HCT116 (B) and SW48 (C) cells. D, immunofluorescent staining of colorectal tumor and paired normal mucosa from patient #2 revealed that β-catenin is highly expressed in the intestinal crypt. E, the hSETD1A complex colocalizes with the β-catenin complex at the known β-catenin responsive element at MYC gene in HCT116 cells. ChIP analysis was performed using antibodies against hSETD1A complex (hSETD1A, Ash2L, WDR5, and RbBP5) and β-catenin complex (β-catenin and TCF4) to determine their binding to the known β-catenin responsive element at MYC enhancer and a negative control region located 70 Kb downstream the TAL1 promoter in HCT116. F and G, H3K4me3 (F), and β-catenin (G) ChIP assays were performed in the Scramble (Scr) control and hSETD1A knockdown HCT116 cells. H3K4me3 enrichment was decreased at the TSSs of the selected Wnt target genes upon hSETD1A knockdown, whereas β-catenin binding remained unchanged. The VEGFA 3′UTR was shown as a negative control region. Data, mean ± SD from three independent replicates. *, P

    Techniques Used: Western Blot, Staining, Chromatin Immunoprecipitation, Binding Assay, Negative Control

    hSETD1A plays an important role in colorectal tumor engraftment. A, ChIP assay revealed that increased hSETD1A levels associate with increased H3K4me3 occupancy at the TSSs of Wnt target genes except for E-cadherin ( CDH1 ), which is repressed by the Wnt pathway in human colorectal cancer compared to adjacent normal mucosa from patient #1. B, Wnt target genes are highly active in human colorectal tumors in which hSETD1A is aberrantly expressed (patients #1–3), but are less active in tumors in which hSETD1A levels are not upregulated (patients #7 and #10). mRNA levels of Wnt target genes in tumors were determined by qRT-PCR and normalized to that of adjacent normal mucosa from each patient. Data, mean ± SD from three independent replicates. *, P
    Figure Legend Snippet: hSETD1A plays an important role in colorectal tumor engraftment. A, ChIP assay revealed that increased hSETD1A levels associate with increased H3K4me3 occupancy at the TSSs of Wnt target genes except for E-cadherin ( CDH1 ), which is repressed by the Wnt pathway in human colorectal cancer compared to adjacent normal mucosa from patient #1. B, Wnt target genes are highly active in human colorectal tumors in which hSETD1A is aberrantly expressed (patients #1–3), but are less active in tumors in which hSETD1A levels are not upregulated (patients #7 and #10). mRNA levels of Wnt target genes in tumors were determined by qRT-PCR and normalized to that of adjacent normal mucosa from each patient. Data, mean ± SD from three independent replicates. *, P

    Techniques Used: Chromatin Immunoprecipitation, Quantitative RT-PCR

    hSETD1A and H3K4me3 are upregulated in human colorectal cancer (CRC). A, hSETD1A and H3K4me3 were upregulated in various human colorectal cancer cell lines compared with normal intestinal epithelial cell line FHs int-74 as detected by Western blot analysis. B, hSETD1A mRNA levels were upregulated in human colorectal tumors compared with adjacent paired normal mucosa by qRT-PCR. Data, mean ± SD from three independent replicates. *, P
    Figure Legend Snippet: hSETD1A and H3K4me3 are upregulated in human colorectal cancer (CRC). A, hSETD1A and H3K4me3 were upregulated in various human colorectal cancer cell lines compared with normal intestinal epithelial cell line FHs int-74 as detected by Western blot analysis. B, hSETD1A mRNA levels were upregulated in human colorectal tumors compared with adjacent paired normal mucosa by qRT-PCR. Data, mean ± SD from three independent replicates. *, P

    Techniques Used: Western Blot, Quantitative RT-PCR

    5) Product Images from "Tumor-derived tissue factor activates coagulation and enhances thrombosis in a mouse xenograft model of human pancreatic cancer"

    Article Title: Tumor-derived tissue factor activates coagulation and enhances thrombosis in a mouse xenograft model of human pancreatic cancer

    Journal: Blood

    doi: 10.1182/blood-2012-01-402156

    Analysis of TF mRNA expression in human pancreatic and colorectal cancer cell lines
    Figure Legend Snippet: Analysis of TF mRNA expression in human pancreatic and colorectal cancer cell lines

    Techniques Used: Expressing

    6) Product Images from "POFUT1 as a Promising Novel Biomarker of Colorectal Cancer"

    Article Title: POFUT1 as a Promising Novel Biomarker of Colorectal Cancer

    Journal: Cancers

    doi: 10.3390/cancers10110411

    POFUT1 expression in colorectal tissues and cell lines. Immunohistochemistry analysis ( A ) shows that POFUT1 is overexpressed in tumor tissues from first colorectal cancer (CRC) stage. Immunofluorescence labeling of POFUT1 (red), performed on wild-type and POFUT1 knockdown HCT 116 and SW620 human colorectal cancer cell lines, confirmed the antibody specificity ( B ). Western blot realized on colorectal tissues ( C ) and colorectal CCD841CoN, HCT 116, HT-29 and SW620 cell lines ( D ) validate the POFUT1 overexpression in cancer samples compared to healthy samples.
    Figure Legend Snippet: POFUT1 expression in colorectal tissues and cell lines. Immunohistochemistry analysis ( A ) shows that POFUT1 is overexpressed in tumor tissues from first colorectal cancer (CRC) stage. Immunofluorescence labeling of POFUT1 (red), performed on wild-type and POFUT1 knockdown HCT 116 and SW620 human colorectal cancer cell lines, confirmed the antibody specificity ( B ). Western blot realized on colorectal tissues ( C ) and colorectal CCD841CoN, HCT 116, HT-29 and SW620 cell lines ( D ) validate the POFUT1 overexpression in cancer samples compared to healthy samples.

    Techniques Used: Expressing, Immunohistochemistry, Immunofluorescence, Labeling, Western Blot, Over Expression

    POFUT1 is overexpressed in most of cancer types especially in colorectal cancer from the first stage. RNAseq data from FireBrowse database show that in 22 cancer types (including COAD and READ), POFUT1 expression is higher than in the corresponding normal tissues and for 6 cancer types it is the reverse ( A ). Data are missing for nine cancer types. ACC: adrenocortical carcinoma, BLCA: bladder urothelial Carcinoma, BRCA: breast invasive carcinoma, CESC: cervical squamous cell carcinoma and endocervical adenocarcinoma, CHOL: cholangiocarcinoma, COAD: colon adenocarcinoma, COADREAD: colorectal adenocarcinoma, DLBC: lymphoid neoplasm diffuse large B-cell lymphoma, ESCA: esophageal carcinoma, GBM: glioblastoma multiforme, GBMLGG: glioma, HNSC: head and neck squamous cell carcinoma, KICH: kidney chromophobe, KIPAN: pan-kidney cohort, KIRC: kidney renal clear cell carcinoma, KIRP: kidney renal papillary cell carcinoma, LAML: acute myeloid leukemia, LGG: brain lower grade glioma, LIHC: liver hepatocellular carcinoma, LUAD: lung adenocarcinoma, LUSC: lung squamous cell carcinoma, MESO: mesothelioma, OV: ovarian serous cystadenocarcinoma, PAAD: pancreatic adenocarcinoma, PCPG: pheochromocytoma and paraganglioma, PRAD: prostate adenocarcinoma, READ: rectum adenocarcinoma, SARC: sarcoma, SKCM: skin Cutaneous Melanoma, STAD: stomach adenocarcinoma, STES: stomach and esophageal carcinoma, TGCT: testicular germ cell tumors, THCA: thyroid carcinoma, THYM: Thymoma, UCEC: uterine corpus endometrial carcinoma, UCS: uterine carcinosarcoma, UVM: uveal melanoma. COADREAD RNAseq data extracted from FireBrowse database containing 626 CRC and 51 normal adjacent tissues show that POFUT1 is significantly overexpressed in tumor tissues ( B ) and from the first stage of tumor classification ( C ). For B. and C., bar graph represented mean of log2 RSEM ± SEM. Statistical significance was assessed using a two-tailed Student test; * p
    Figure Legend Snippet: POFUT1 is overexpressed in most of cancer types especially in colorectal cancer from the first stage. RNAseq data from FireBrowse database show that in 22 cancer types (including COAD and READ), POFUT1 expression is higher than in the corresponding normal tissues and for 6 cancer types it is the reverse ( A ). Data are missing for nine cancer types. ACC: adrenocortical carcinoma, BLCA: bladder urothelial Carcinoma, BRCA: breast invasive carcinoma, CESC: cervical squamous cell carcinoma and endocervical adenocarcinoma, CHOL: cholangiocarcinoma, COAD: colon adenocarcinoma, COADREAD: colorectal adenocarcinoma, DLBC: lymphoid neoplasm diffuse large B-cell lymphoma, ESCA: esophageal carcinoma, GBM: glioblastoma multiforme, GBMLGG: glioma, HNSC: head and neck squamous cell carcinoma, KICH: kidney chromophobe, KIPAN: pan-kidney cohort, KIRC: kidney renal clear cell carcinoma, KIRP: kidney renal papillary cell carcinoma, LAML: acute myeloid leukemia, LGG: brain lower grade glioma, LIHC: liver hepatocellular carcinoma, LUAD: lung adenocarcinoma, LUSC: lung squamous cell carcinoma, MESO: mesothelioma, OV: ovarian serous cystadenocarcinoma, PAAD: pancreatic adenocarcinoma, PCPG: pheochromocytoma and paraganglioma, PRAD: prostate adenocarcinoma, READ: rectum adenocarcinoma, SARC: sarcoma, SKCM: skin Cutaneous Melanoma, STAD: stomach adenocarcinoma, STES: stomach and esophageal carcinoma, TGCT: testicular germ cell tumors, THCA: thyroid carcinoma, THYM: Thymoma, UCEC: uterine corpus endometrial carcinoma, UCS: uterine carcinosarcoma, UVM: uveal melanoma. COADREAD RNAseq data extracted from FireBrowse database containing 626 CRC and 51 normal adjacent tissues show that POFUT1 is significantly overexpressed in tumor tissues ( B ) and from the first stage of tumor classification ( C ). For B. and C., bar graph represented mean of log2 RSEM ± SEM. Statistical significance was assessed using a two-tailed Student test; * p

    Techniques Used: Expressing, Two Tailed Test

    7) Product Images from "TCF4 Is a Molecular Target of Resveratrol in the Prevention of Colorectal Cancer"

    Article Title: TCF4 Is a Molecular Target of Resveratrol in the Prevention of Colorectal Cancer

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms160510411

    T-cell factor 4 (TCF4) is a potential molecular target of phytochemicals in HCT116 cells. ( A ) Normal human colon cells and different types of human colorectal cancer cells (HCT116, SW480, LoVo and Caco-2) were lysed, and Western blot was performed for TCF4, β-catenin and actin, as described in the Experimental Section; ( B ) Normal human colon cells and different types of human colorectal cancer cells (HCT116, SW480, HT-29, LoVo and Caco-2) were grown overnight. Total RNA was extracted, and RT-PCR was performed using primers for TCF4 and GAPDH; ( C ) HCT116 cells were treated with 50 µM of epigallocatechin gallate (EGCG), resveratrol, genistein and capsaicin for 24 h, and a Western blot was performed for TCF4, β-catenin and actin.
    Figure Legend Snippet: T-cell factor 4 (TCF4) is a potential molecular target of phytochemicals in HCT116 cells. ( A ) Normal human colon cells and different types of human colorectal cancer cells (HCT116, SW480, LoVo and Caco-2) were lysed, and Western blot was performed for TCF4, β-catenin and actin, as described in the Experimental Section; ( B ) Normal human colon cells and different types of human colorectal cancer cells (HCT116, SW480, HT-29, LoVo and Caco-2) were grown overnight. Total RNA was extracted, and RT-PCR was performed using primers for TCF4 and GAPDH; ( C ) HCT116 cells were treated with 50 µM of epigallocatechin gallate (EGCG), resveratrol, genistein and capsaicin for 24 h, and a Western blot was performed for TCF4, β-catenin and actin.

    Techniques Used: Western Blot, Reverse Transcription Polymerase Chain Reaction

    8) Product Images from "Anti-helminth compound niclosamide downregulates Wnt Signaling and elicits antitumor responses in tumors with activating APC mutations"

    Article Title: Anti-helminth compound niclosamide downregulates Wnt Signaling and elicits antitumor responses in tumors with activating APC mutations

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-10-3978

    Niclosamide inhibits proliferation of colorectal cancer cell lines and explants
    Figure Legend Snippet: Niclosamide inhibits proliferation of colorectal cancer cell lines and explants

    Techniques Used:

    Niclosamide inhibits Wnt/β-catenin signaling and downregulates dishevelled 2 and β-catenin expression by colorectal cancer cells in vitro
    Figure Legend Snippet: Niclosamide inhibits Wnt/β-catenin signaling and downregulates dishevelled 2 and β-catenin expression by colorectal cancer cells in vitro

    Techniques Used: Expressing, In Vitro

    Combination effect of niclosamide and oxaliplatin on colorectal cancer cell lines and explants
    Figure Legend Snippet: Combination effect of niclosamide and oxaliplatin on colorectal cancer cell lines and explants

    Techniques Used:

    Niclosamide inhibits the growth of colorectal cancers in NOD/SCID mice and downregulates dishevelled 2 and β-catenin expression
    Figure Legend Snippet: Niclosamide inhibits the growth of colorectal cancers in NOD/SCID mice and downregulates dishevelled 2 and β-catenin expression

    Techniques Used: Mouse Assay, Expressing

    Niclosamide inhibits the proliferation of colorectal cancer cell Lines and has minimal toxicity on normal fibroblasts, PBMCs, or immortalized mammary epithelial cells
    Figure Legend Snippet: Niclosamide inhibits the proliferation of colorectal cancer cell Lines and has minimal toxicity on normal fibroblasts, PBMCs, or immortalized mammary epithelial cells

    Techniques Used:

    9) Product Images from "CDX2 is an amplified lineage-survival oncogene in colorectal cancer"

    Article Title: CDX2 is an amplified lineage-survival oncogene in colorectal cancer

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1206004109

    CDX2 is frequently amplified in colorectal cancer, where it promotes Wnt/β-catenin signaling. ( Upper ) Plot of significant DNA amplifications in colorectal cancers reveals amplification on chromosome 13 pinpointing CDX2 . ( Lower ) Schematic model
    Figure Legend Snippet: CDX2 is frequently amplified in colorectal cancer, where it promotes Wnt/β-catenin signaling. ( Upper ) Plot of significant DNA amplifications in colorectal cancers reveals amplification on chromosome 13 pinpointing CDX2 . ( Lower ) Schematic model

    Techniques Used: Amplification

    Recurrent amplification and overexpression of CDX2 in colorectal cancer cell lines and primary tumors. ( A ) Peaks of recurrent amplification among 29 colorectal cancer cell lines. The G-score is shown on the upper x -axis, and the q -value is shown on the
    Figure Legend Snippet: Recurrent amplification and overexpression of CDX2 in colorectal cancer cell lines and primary tumors. ( A ) Peaks of recurrent amplification among 29 colorectal cancer cell lines. The G-score is shown on the upper x -axis, and the q -value is shown on the

    Techniques Used: Amplification, Over Expression

    Recurrent 13q12 Amplification in Colorectal Cancer Targets CDX2 .
    Figure Legend Snippet: Recurrent 13q12 Amplification in Colorectal Cancer Targets CDX2 .

    Techniques Used: Amplification

    10) Product Images from "Xanthohumol suppresses oestrogen-signalling in breast cancer through the inhibition of BIG3-PHB2 interactions"

    Article Title: Xanthohumol suppresses oestrogen-signalling in breast cancer through the inhibition of BIG3-PHB2 interactions

    Journal: Scientific Reports

    doi: 10.1038/srep07355

    Xanthohumol suppresses E2-dependent cell growth independently of apoptosis induction and VCP function. (a) Flow cytometric analyses were performed to evaluate the effect of XN treatment on the cell cycle. MCF-7 cells were treated for 24 h with E2 and/or XN or ERAP. (b) Representative cell morphologies of MCF-7 or MCF-10A cell following XN treatment are shown. (c) Expression patterns of VCP, BIG3, and PHB2 in breast cancer and normal epithelial cell lines. Colorectal cancer cell lines (HCT116 and SW480) and an epidermoid carcinoma cell line (A431) were used as positive controls for VCP expression. β-Actin served as a quantitative internal control. The blots were cropped, and the full-length blots were included in the supplementary information. (d) The CHOP and GRP78 expression levels following XN treatment were evaluated using real-time PCR. The data represent the mean ± SE of three independent experiments (* P
    Figure Legend Snippet: Xanthohumol suppresses E2-dependent cell growth independently of apoptosis induction and VCP function. (a) Flow cytometric analyses were performed to evaluate the effect of XN treatment on the cell cycle. MCF-7 cells were treated for 24 h with E2 and/or XN or ERAP. (b) Representative cell morphologies of MCF-7 or MCF-10A cell following XN treatment are shown. (c) Expression patterns of VCP, BIG3, and PHB2 in breast cancer and normal epithelial cell lines. Colorectal cancer cell lines (HCT116 and SW480) and an epidermoid carcinoma cell line (A431) were used as positive controls for VCP expression. β-Actin served as a quantitative internal control. The blots were cropped, and the full-length blots were included in the supplementary information. (d) The CHOP and GRP78 expression levels following XN treatment were evaluated using real-time PCR. The data represent the mean ± SE of three independent experiments (* P

    Techniques Used: Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction

    11) Product Images from "Therapeutic potential of targeting S100A11 in malignant pleural mesothelioma"

    Article Title: Therapeutic potential of targeting S100A11 in malignant pleural mesothelioma

    Journal: Oncogenesis

    doi: 10.1038/s41389-017-0017-3

    High secretion and overexpression of S100A11 in MPM. a Concentration of secreted S100A11 in the culture media of various cell lines, as determined by ELISA. The secretion level of S100A11 in all MPM cell lines, except for 211H, is higher than that of NM. 211H MSTO-211H, NM normal mesothelial cell, LC lung cancer, GC gastric cancer, CRC colorectal cancer, BC breast cancer. b Protein level of S100A11 was higher in MPM cell lines, compared to NM. NM normal mesothelial cell. c Immunohistochemical analysis of S100A11 in surgically resected tissues from patients with MPM. The representative images of three MPM subtypes are shown. The pictures on the left show hematoxylin–eosin (HE)-stained images corresponding to the S100A11-stained images in the middle. Scale bars, 50 µm (positive and negative control) and 100 µm (clinical samples)
    Figure Legend Snippet: High secretion and overexpression of S100A11 in MPM. a Concentration of secreted S100A11 in the culture media of various cell lines, as determined by ELISA. The secretion level of S100A11 in all MPM cell lines, except for 211H, is higher than that of NM. 211H MSTO-211H, NM normal mesothelial cell, LC lung cancer, GC gastric cancer, CRC colorectal cancer, BC breast cancer. b Protein level of S100A11 was higher in MPM cell lines, compared to NM. NM normal mesothelial cell. c Immunohistochemical analysis of S100A11 in surgically resected tissues from patients with MPM. The representative images of three MPM subtypes are shown. The pictures on the left show hematoxylin–eosin (HE)-stained images corresponding to the S100A11-stained images in the middle. Scale bars, 50 µm (positive and negative control) and 100 µm (clinical samples)

    Techniques Used: Over Expression, Concentration Assay, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining, Negative Control

    12) Product Images from "Vulnerability to low-dose combination of irinotecan and niraparib in ATM-mutated colorectal cancer"

    Article Title: Vulnerability to low-dose combination of irinotecan and niraparib in ATM-mutated colorectal cancer

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/s13046-020-01811-8

    Ex vivo testing of niraparib, SN38, and their combination on human primary CRC 3D cell cultures (spheroids) according to the genetic background. Five primary 3D cell cultures (spheroids) were generated from surgical specimens (primary tumor or metastasis) of colorectal cancer. Tumor specimens were genetically profiled using a comprehensive genomic panel encompassing 324 frequently mutated cancer genes. Growth inhibition is significantly improved in IC-006 (ATM-mutated) at two concentration levels and in IC-011 (CHEK2-mutated). * p
    Figure Legend Snippet: Ex vivo testing of niraparib, SN38, and their combination on human primary CRC 3D cell cultures (spheroids) according to the genetic background. Five primary 3D cell cultures (spheroids) were generated from surgical specimens (primary tumor or metastasis) of colorectal cancer. Tumor specimens were genetically profiled using a comprehensive genomic panel encompassing 324 frequently mutated cancer genes. Growth inhibition is significantly improved in IC-006 (ATM-mutated) at two concentration levels and in IC-011 (CHEK2-mutated). * p

    Techniques Used: Ex Vivo, Generated, Inhibition, Concentration Assay

    13) Product Images from "Fusobacterium nucleatum Promotes the Progression of Colorectal Cancer Through Cdk5-Activated Wnt/β-Catenin Signaling"

    Article Title: Fusobacterium nucleatum Promotes the Progression of Colorectal Cancer Through Cdk5-Activated Wnt/β-Catenin Signaling

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2020.545251

    Upregulation of cyclin-dependent kinase 5 (Cdk5) protein expression and its correlation with Fusobacterium nucleatum in colorectal cancer (CRC). (A) Cdk5 protein levels were detected by Western blot (WB) analysis in CRC and normal tissues. (B) Statistical analysis of Cdk5 mRNA expression in (A) (* P
    Figure Legend Snippet: Upregulation of cyclin-dependent kinase 5 (Cdk5) protein expression and its correlation with Fusobacterium nucleatum in colorectal cancer (CRC). (A) Cdk5 protein levels were detected by Western blot (WB) analysis in CRC and normal tissues. (B) Statistical analysis of Cdk5 mRNA expression in (A) (* P

    Techniques Used: Expressing, Western Blot

    14) Product Images from "circREEP3 Drives Colorectal Cancer Progression via Activation of FKBP10 Transcription and Restriction of Antitumor Immunity, circREEP3 Drives Colorectal Cancer Progression via Activation of FKBP10 Transcription and Restriction of Antitumor Immunity"

    Article Title: circREEP3 Drives Colorectal Cancer Progression via Activation of FKBP10 Transcription and Restriction of Antitumor Immunity, circREEP3 Drives Colorectal Cancer Progression via Activation of FKBP10 Transcription and Restriction of Antitumor Immunity

    Journal: Advanced Science

    doi: 10.1002/advs.202105160

    circREEP3 promotes CRC progression via FKBP10. A) FKBP10 expression intensity was analyzed according to online data sets (GSE44076, GSE44861, and GSE68486). Healthy, healthy colon mucosa; normal, normal distant colon mucosa from tumor patients; tumor, colon cancer tissues. B) Western blotting analysis of FKBP10 expression in CRC tissues and paired adjacent normal tissues. C) FKBP10 expression was analyzed by IHC staining in paired CRC tissues and normal tissues. Scale bar, 100 µm. D) Survival rate was analyzed based on FKBP10 expression according to online data set GSE17536. E) Survival rate was analyzed by Kaplan–Meier method. CRC array tissues were divided into FKBP10 high and low subgroups based on FKBP10 expression. F,G) CCK8 and colony formation assays were conducted using indicated LoVo and HCT116 cell lines to test cell proliferation. Ctrl, control. H) Transwell assay was carried out to evaluate the invasion potential of LoVo and HCT116 cell lines. I) Indicated LoVo cells were used for xenograft assay. Tumor volumes were measured every 5 d. n = 5 for each group. J) Tumor weights were determined on day 25 postinjection. n = 5 for each group. K) Potential of lung metastasis was determined via vein tail injection of indicated LoVo cells. The number of metastatic nodules in the lung was calculated in the right panel. n = 5 mice each group. * P
    Figure Legend Snippet: circREEP3 promotes CRC progression via FKBP10. A) FKBP10 expression intensity was analyzed according to online data sets (GSE44076, GSE44861, and GSE68486). Healthy, healthy colon mucosa; normal, normal distant colon mucosa from tumor patients; tumor, colon cancer tissues. B) Western blotting analysis of FKBP10 expression in CRC tissues and paired adjacent normal tissues. C) FKBP10 expression was analyzed by IHC staining in paired CRC tissues and normal tissues. Scale bar, 100 µm. D) Survival rate was analyzed based on FKBP10 expression according to online data set GSE17536. E) Survival rate was analyzed by Kaplan–Meier method. CRC array tissues were divided into FKBP10 high and low subgroups based on FKBP10 expression. F,G) CCK8 and colony formation assays were conducted using indicated LoVo and HCT116 cell lines to test cell proliferation. Ctrl, control. H) Transwell assay was carried out to evaluate the invasion potential of LoVo and HCT116 cell lines. I) Indicated LoVo cells were used for xenograft assay. Tumor volumes were measured every 5 d. n = 5 for each group. J) Tumor weights were determined on day 25 postinjection. n = 5 for each group. K) Potential of lung metastasis was determined via vein tail injection of indicated LoVo cells. The number of metastatic nodules in the lung was calculated in the right panel. n = 5 mice each group. * P

    Techniques Used: Expressing, Western Blot, Immunohistochemistry, Staining, Transwell Assay, Xenograft Assay, Injection, Mouse Assay

    circREEP3 interacts with CHD7 to activate FKBP10 transcription. A) CRC sample cells were lysed and incubated with biotin‐labeled and linearized circREEP3 transcripts, anti‐sense or beads control. Precipitants were resolved by SDS‐PAGE, followed by silver staining. Indicated bands were identified via mass spectrometry. B) RNA pulldown was performed using CRC sample cell lysates and biotin‐labeled circREEP3 . Then precipitates were detected using anti‐CHD7 by Western blotting (upper panels) and total CHD7 levels were also examined (lower panel). C) RIP (RNA Immunoprecipitation) assay was performed using anti‐CHD7 to detect the interaction between circREEP3 and CHD7 in CRC cells. D) Electrophoretic mobility shift assay (EMSA) using biotin‐labeled circREEP3 and nuclear extract with or without anti‐CHD7. E) circREEP3 was colocalized with CHD7 in CRC cells by immunofluorescence staining. Scale bar, 10 µm. F) RNA pulldown assay was conducted using WT circREEP3 and its different mutations. HR, hairpin region. The CHD7 levels in circRNA pulldown eluate (upper panel) and the input sample (lower panel) were shown. G) EMSA assay using biotin‐labeled WT circREEP3 or indicated mutation with or without anti‐CHD7. H) ChIP assay was performed to test the enrichment of CHD7 on FKBP10 promoter. I) ChIP assay was conducted to measure CHD7 enrichment on FKBP10 promoter in circREEP3 +/+ or circREEP3 −/− LoVo cells. J) Western blot of CHD7 in eluate from CAPTURE assay (CRISPR affinity purification in situ of regulatory elements) using sgRNA (small guide RNA) targeting FKBP10 promoter. Pro, promoter. K) DNaseI accessibility assay was conducted using indicated LoVo and HCT116 cells. n = 3 independent samples. L) ChIP assay was performed to measure the enrichment of RNA Pol II on FKBP10 promoter in indicated LoVo and HCT116 cells. M) Indicated LoVo and HCT116 cells were subjected to nuclear run‐on assay, followed by detection of FKBP10 transcription through qPCR analysis. n = 3 independent samples. N) Western blotting was performed to test FKBP10 expression in indicated LoVo and HCT116 cells. * P
    Figure Legend Snippet: circREEP3 interacts with CHD7 to activate FKBP10 transcription. A) CRC sample cells were lysed and incubated with biotin‐labeled and linearized circREEP3 transcripts, anti‐sense or beads control. Precipitants were resolved by SDS‐PAGE, followed by silver staining. Indicated bands were identified via mass spectrometry. B) RNA pulldown was performed using CRC sample cell lysates and biotin‐labeled circREEP3 . Then precipitates were detected using anti‐CHD7 by Western blotting (upper panels) and total CHD7 levels were also examined (lower panel). C) RIP (RNA Immunoprecipitation) assay was performed using anti‐CHD7 to detect the interaction between circREEP3 and CHD7 in CRC cells. D) Electrophoretic mobility shift assay (EMSA) using biotin‐labeled circREEP3 and nuclear extract with or without anti‐CHD7. E) circREEP3 was colocalized with CHD7 in CRC cells by immunofluorescence staining. Scale bar, 10 µm. F) RNA pulldown assay was conducted using WT circREEP3 and its different mutations. HR, hairpin region. The CHD7 levels in circRNA pulldown eluate (upper panel) and the input sample (lower panel) were shown. G) EMSA assay using biotin‐labeled WT circREEP3 or indicated mutation with or without anti‐CHD7. H) ChIP assay was performed to test the enrichment of CHD7 on FKBP10 promoter. I) ChIP assay was conducted to measure CHD7 enrichment on FKBP10 promoter in circREEP3 +/+ or circREEP3 −/− LoVo cells. J) Western blot of CHD7 in eluate from CAPTURE assay (CRISPR affinity purification in situ of regulatory elements) using sgRNA (small guide RNA) targeting FKBP10 promoter. Pro, promoter. K) DNaseI accessibility assay was conducted using indicated LoVo and HCT116 cells. n = 3 independent samples. L) ChIP assay was performed to measure the enrichment of RNA Pol II on FKBP10 promoter in indicated LoVo and HCT116 cells. M) Indicated LoVo and HCT116 cells were subjected to nuclear run‐on assay, followed by detection of FKBP10 transcription through qPCR analysis. n = 3 independent samples. N) Western blotting was performed to test FKBP10 expression in indicated LoVo and HCT116 cells. * P

    Techniques Used: Incubation, Labeling, SDS Page, Silver Staining, Mass Spectrometry, Western Blot, Immunoprecipitation, Electrophoretic Mobility Shift Assay, Immunofluorescence, Staining, Mutagenesis, Chromatin Immunoprecipitation, CRISPR, Affinity Purification, In Situ, Nuclear Run-on Assay, Real-time Polymerase Chain Reaction, Expressing

    circREEP3 contributes to CRC stem cell phenotype. A) CRC sample cells were sorted by gating on CD133 and divided into CD133 high (CD133 high ) and low (CD133 low ) subgroups. Then Northern blotting was performed to detect circREEP3 expression. 18S was the loading control. B) CRC sample cells were used for sphere formation. Then circREEP3 levels were measured by Northern blotting. 18S was the loading control. C) CRC sample cells with circREEP3 silencing were used for sphere formation, followed by detection of CD133 expression through FACS analysis. shCtrl, control shRNA. D) circREEP3 knockdown and control CRC sample cells were used for sphere formation, followed by detection of CD133 expression through FACS analysis. Vec, empty vector. E,F) Sphere formation assay using circREEP3 ‐silenced or overexpressing CRC cells. G,H) Tumor initiation assay using circREEP3 silenced or overexpressing CRC sample cells were conducted and the ratios of tumor‐free mice were calculated. n = 10 for each group. * P
    Figure Legend Snippet: circREEP3 contributes to CRC stem cell phenotype. A) CRC sample cells were sorted by gating on CD133 and divided into CD133 high (CD133 high ) and low (CD133 low ) subgroups. Then Northern blotting was performed to detect circREEP3 expression. 18S was the loading control. B) CRC sample cells were used for sphere formation. Then circREEP3 levels were measured by Northern blotting. 18S was the loading control. C) CRC sample cells with circREEP3 silencing were used for sphere formation, followed by detection of CD133 expression through FACS analysis. shCtrl, control shRNA. D) circREEP3 knockdown and control CRC sample cells were used for sphere formation, followed by detection of CD133 expression through FACS analysis. Vec, empty vector. E,F) Sphere formation assay using circREEP3 ‐silenced or overexpressing CRC cells. G,H) Tumor initiation assay using circREEP3 silenced or overexpressing CRC sample cells were conducted and the ratios of tumor‐free mice were calculated. n = 10 for each group. * P

    Techniques Used: Northern Blot, Expressing, FACS, shRNA, Plasmid Preparation, Tube Formation Assay, Mouse Assay

    circREEP3 promotes CRC growth and metastasis. A) Relative expression of circREEP3 in CRC cell lines was measured by qPCR. B) CCK8 assay for analysis of cell proliferation ability in circREEP3 +/+ or circREEP3 − /− LoVo and HCT116 cells. C) CCK8 assay for analysis of cell proliferation ability in circREEP3 ‐overexpressing (oe) or control LoVo and HCT116 cells. Vec, empty vector. D) Colony formation assay was conducted to test cell proliferation using circREEP3 +/+ or circREEP3 −/− LoVo and HCT116 cells. E) Colony formation assay was conducted to test cell proliferation using circREEP3 ‐overexpressing or control LoVo and HCT116 cells. F) Transwell assay using circREEP3 +/+ or circREEP3 −/− LoVo and HCT116 cells was carried out. Invaded cells were calculated. G) Transwell assay using circREEP3 ‐overexpressing or control LoVo and HCT116 cells was carried out. H) circREEP3 +/+ or circREEP3 −/− LoVo cells were used for xenograft assay. Tumor volumes were measured every 5 d. n = 5 for each group. I) Tumor weights were determined on day 25 postinjection. Typical luciferase images of circREEP3 +/+ or circREEP3 −/− tumors were in the right. n = 5 for each group. J) Immunohistochemistry (IHC) analysis for Ki67 expression in tumor tissues of (I). Percentage of Ki67 + cells was calculated and shown in the right panel. Scale bar, 100 µm. K) circREEP3 highly ( circREEP3 high ) or lowly ( circREEP3 low ) expressed CRC tumor cells were used for xenograft assay through NSG (NOD/Scid/Il2rg) mice. Tumor volumes were calculated every 5 d. L) Potential of lung metastasis was measured through vein tail injection of circREEP3 +/+ or circREEP3 −/− LoVo cells. Representative images were shown in the left panel. The number of metastatic nodules in the lung was calculated and presented in the right panel. n = 5 mice each group. w, week. * P
    Figure Legend Snippet: circREEP3 promotes CRC growth and metastasis. A) Relative expression of circREEP3 in CRC cell lines was measured by qPCR. B) CCK8 assay for analysis of cell proliferation ability in circREEP3 +/+ or circREEP3 − /− LoVo and HCT116 cells. C) CCK8 assay for analysis of cell proliferation ability in circREEP3 ‐overexpressing (oe) or control LoVo and HCT116 cells. Vec, empty vector. D) Colony formation assay was conducted to test cell proliferation using circREEP3 +/+ or circREEP3 −/− LoVo and HCT116 cells. E) Colony formation assay was conducted to test cell proliferation using circREEP3 ‐overexpressing or control LoVo and HCT116 cells. F) Transwell assay using circREEP3 +/+ or circREEP3 −/− LoVo and HCT116 cells was carried out. Invaded cells were calculated. G) Transwell assay using circREEP3 ‐overexpressing or control LoVo and HCT116 cells was carried out. H) circREEP3 +/+ or circREEP3 −/− LoVo cells were used for xenograft assay. Tumor volumes were measured every 5 d. n = 5 for each group. I) Tumor weights were determined on day 25 postinjection. Typical luciferase images of circREEP3 +/+ or circREEP3 −/− tumors were in the right. n = 5 for each group. J) Immunohistochemistry (IHC) analysis for Ki67 expression in tumor tissues of (I). Percentage of Ki67 + cells was calculated and shown in the right panel. Scale bar, 100 µm. K) circREEP3 highly ( circREEP3 high ) or lowly ( circREEP3 low ) expressed CRC tumor cells were used for xenograft assay through NSG (NOD/Scid/Il2rg) mice. Tumor volumes were calculated every 5 d. L) Potential of lung metastasis was measured through vein tail injection of circREEP3 +/+ or circREEP3 −/− LoVo cells. Representative images were shown in the left panel. The number of metastatic nodules in the lung was calculated and presented in the right panel. n = 5 mice each group. w, week. * P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, CCK-8 Assay, Plasmid Preparation, Colony Assay, Transwell Assay, Xenograft Assay, Luciferase, Immunohistochemistry, Mouse Assay, Injection

    circREEP3 initiates FKBP10 transcription. A) Differentially expressed genes in circREEP3 +/+ or circREEP3 −/− LoVo cells. B) qPCR validation of the most downregulated or upregulated genes in circREEP3 −/− LoVo cells. C) Western blotting analysis for FKBP10 expression in circREEP3 −/− LoVo cells. D) ChIRP (Chromatin Isolation by RNA Purification) assay was performed using biotin‐labeled circREEP3 probes and enrichment of circREEP3 in FKBP10 promoter was analyzed. E) circREEP3 linearized RNA was immobilized on NC membranes, followed by probing with indicated biotin‐labeled DNA probes. F) Luciferase reporter assay using FKBP10 truncated promoters with overexpression of circREEP3 or vector control. G,H) ChIP assay was carried out to analyze the enrichment of H3K27ac or H3K27me3 on FKBP10 promoter. I) DNaseI accessibility assay was conducted using circREEP3 +/+ or circREEP3 −/− LoVo cells. n = 3 independent samples. J) ChIP assay was conducted to detect the enrichment of RNA pol II Ser2P on FKBP10 promoter. K) circREEP3 +/+ or circREEP3 −/− LoVo cells were subjected to nuclear run‐on assay, followed by detection of FKBP10 transcription through qPCR analysis. n = 3 independent samples. * P
    Figure Legend Snippet: circREEP3 initiates FKBP10 transcription. A) Differentially expressed genes in circREEP3 +/+ or circREEP3 −/− LoVo cells. B) qPCR validation of the most downregulated or upregulated genes in circREEP3 −/− LoVo cells. C) Western blotting analysis for FKBP10 expression in circREEP3 −/− LoVo cells. D) ChIRP (Chromatin Isolation by RNA Purification) assay was performed using biotin‐labeled circREEP3 probes and enrichment of circREEP3 in FKBP10 promoter was analyzed. E) circREEP3 linearized RNA was immobilized on NC membranes, followed by probing with indicated biotin‐labeled DNA probes. F) Luciferase reporter assay using FKBP10 truncated promoters with overexpression of circREEP3 or vector control. G,H) ChIP assay was carried out to analyze the enrichment of H3K27ac or H3K27me3 on FKBP10 promoter. I) DNaseI accessibility assay was conducted using circREEP3 +/+ or circREEP3 −/− LoVo cells. n = 3 independent samples. J) ChIP assay was conducted to detect the enrichment of RNA pol II Ser2P on FKBP10 promoter. K) circREEP3 +/+ or circREEP3 −/− LoVo cells were subjected to nuclear run‐on assay, followed by detection of FKBP10 transcription through qPCR analysis. n = 3 independent samples. * P

    Techniques Used: Real-time Polymerase Chain Reaction, Western Blot, Expressing, Isolation, Purification, Labeling, Luciferase, Reporter Assay, Over Expression, Plasmid Preparation, Chromatin Immunoprecipitation, Nuclear Run-on Assay

    circREEP3 is highly expressed in CRC. A) The most differentially expressed circRNAs in invasive CRC cells compared to those in noninvasive tumor cells were presented. Transwell assay was performed using CRC sample cells. The cells in the lower chamber were considered as invasive tumor cells while the cells in the upper chamber were considered as noninvasive cells. B) The ten most highly expressed circRNAs in invasive CRC cells were selected and their expression levels were validated through qRT‐PCR. C) CCK8 assay was performed to evaluate cell proliferation ability after circRNA knockdown in CRC sample cells. D) Transwell assay was carried out to examine invasion potential after circRNA silencing in CRC tumor cells. E) circREEP3 expression in paired CRC tumor tissues and adjacent normal tissues was measured by Northern blotting. 18S acted as the loading control. F) circREEP3 levels were analyzed in CRC tissue array containing 79 peri‐tumor, 5 stage I, 56 stage II, 39 stage III/IV tissues by in situ hybridization. Typical images containing global views and magnified views were presented in the left panel. Ratios of circRNA positive cells in each sample was calculated and shown in the right panel. Scale bar, 100 µm. G) circREEP3 levels in CRC tissues with distant metastasis or not were measured by in situ hybridization. H) Survival rate was analyzed by Kaplan–Meier method. Samples were divided into circREEP3 high ( circREEP + ≤ 20%) and low ( circREEP + > 20%) subgroups based on circREEP3 median value (20%). I) Fluorescence in situ hybridization was used to test circREEP3 expression in RNase‐R treated CRC sample cells. Scale bar, 10 µm. J) Subcellular location of circRNA in CRC sample cells was analyzed by qPCR. K) RNA abundance was evaluated after treatment with Actinomycin D (Act D, 2 µg mL −1 ). * P
    Figure Legend Snippet: circREEP3 is highly expressed in CRC. A) The most differentially expressed circRNAs in invasive CRC cells compared to those in noninvasive tumor cells were presented. Transwell assay was performed using CRC sample cells. The cells in the lower chamber were considered as invasive tumor cells while the cells in the upper chamber were considered as noninvasive cells. B) The ten most highly expressed circRNAs in invasive CRC cells were selected and their expression levels were validated through qRT‐PCR. C) CCK8 assay was performed to evaluate cell proliferation ability after circRNA knockdown in CRC sample cells. D) Transwell assay was carried out to examine invasion potential after circRNA silencing in CRC tumor cells. E) circREEP3 expression in paired CRC tumor tissues and adjacent normal tissues was measured by Northern blotting. 18S acted as the loading control. F) circREEP3 levels were analyzed in CRC tissue array containing 79 peri‐tumor, 5 stage I, 56 stage II, 39 stage III/IV tissues by in situ hybridization. Typical images containing global views and magnified views were presented in the left panel. Ratios of circRNA positive cells in each sample was calculated and shown in the right panel. Scale bar, 100 µm. G) circREEP3 levels in CRC tissues with distant metastasis or not were measured by in situ hybridization. H) Survival rate was analyzed by Kaplan–Meier method. Samples were divided into circREEP3 high ( circREEP + ≤ 20%) and low ( circREEP + > 20%) subgroups based on circREEP3 median value (20%). I) Fluorescence in situ hybridization was used to test circREEP3 expression in RNase‐R treated CRC sample cells. Scale bar, 10 µm. J) Subcellular location of circRNA in CRC sample cells was analyzed by qPCR. K) RNA abundance was evaluated after treatment with Actinomycin D (Act D, 2 µg mL −1 ). * P

    Techniques Used: Transwell Assay, Expressing, Quantitative RT-PCR, CCK-8 Assay, Northern Blot, In Situ Hybridization, Fluorescence, Real-time Polymerase Chain Reaction

    circREEP3 restricts antitumor immunity via suppression of RIG‐1 signaling. A) GO analysis of the differentially expressed genes in circREEP3 +/+ or circREEP3 −/− LoVo cells. B) Heatmap of RIG‐1 signaling‐related genes according to the RNA sequencing results. C) RNA pulldown was performed using CRC sample cell lysates and biotin‐labeled circREEP3 . Then precipitates were detected using anti‐RIG‐1 by Western blotting. D) RIP assay was performed using anti‐RIG‐1 to detect the interaction between circREEP3 and RIG‐1 in CRC cells. E) Relative expression of RIG‐1 signaling‐related genes was measured in CRC cells by qRT‐PCR. F) Western blot analysis of RIG‐1 protein levels in circREEP3 ‐silenced or control CRC cells. G) Western blot analysis of RIG‐1 protein levels in circREEP3 +/+ or circREEP3 −/− LoVo cells transfected with Flag‐RIG‐1 vector. 36 h after transfection, the cells were treated with cycloheximide (Chx; final concentration: 50 µg mL −1 ). H) Co‐IP assay were performed using anti‐Myc and 293T lysates after transfected with Flag‐RIG‐1, Myc‐RNF125, and circREEP3 . I) Ubiquitination signaling of RIG‐1 was measured by western blot. Indicated plasmids were transfected into 293T cells, followed by MG132 treatment. J) Tumor volumes were measured at indicated time points ( n = 5 mice per group). MC38 cells (5 × 10 5 ) were injected subcutaneously into the right flank of C57BL/6 mice. n = 5 for each group. K) IFN‐ β protein level in the serum was measured by ELISA. L) CD3 + CD8 + T cells in xenograft tumors were detected by FACS. n = 5 tumors per group. ** P
    Figure Legend Snippet: circREEP3 restricts antitumor immunity via suppression of RIG‐1 signaling. A) GO analysis of the differentially expressed genes in circREEP3 +/+ or circREEP3 −/− LoVo cells. B) Heatmap of RIG‐1 signaling‐related genes according to the RNA sequencing results. C) RNA pulldown was performed using CRC sample cell lysates and biotin‐labeled circREEP3 . Then precipitates were detected using anti‐RIG‐1 by Western blotting. D) RIP assay was performed using anti‐RIG‐1 to detect the interaction between circREEP3 and RIG‐1 in CRC cells. E) Relative expression of RIG‐1 signaling‐related genes was measured in CRC cells by qRT‐PCR. F) Western blot analysis of RIG‐1 protein levels in circREEP3 ‐silenced or control CRC cells. G) Western blot analysis of RIG‐1 protein levels in circREEP3 +/+ or circREEP3 −/− LoVo cells transfected with Flag‐RIG‐1 vector. 36 h after transfection, the cells were treated with cycloheximide (Chx; final concentration: 50 µg mL −1 ). H) Co‐IP assay were performed using anti‐Myc and 293T lysates after transfected with Flag‐RIG‐1, Myc‐RNF125, and circREEP3 . I) Ubiquitination signaling of RIG‐1 was measured by western blot. Indicated plasmids were transfected into 293T cells, followed by MG132 treatment. J) Tumor volumes were measured at indicated time points ( n = 5 mice per group). MC38 cells (5 × 10 5 ) were injected subcutaneously into the right flank of C57BL/6 mice. n = 5 for each group. K) IFN‐ β protein level in the serum was measured by ELISA. L) CD3 + CD8 + T cells in xenograft tumors were detected by FACS. n = 5 tumors per group. ** P

    Techniques Used: RNA Sequencing Assay, Labeling, Western Blot, Expressing, Quantitative RT-PCR, Transfection, Plasmid Preparation, Concentration Assay, Co-Immunoprecipitation Assay, Mouse Assay, Injection, Enzyme-linked Immunosorbent Assay, FACS

    15) Product Images from "Transcription factor HOXC10 activates the expression of MTFR2 to regulate the proliferation, invasion and migration of colorectal cancer cells"

    Article Title: Transcription factor HOXC10 activates the expression of MTFR2 to regulate the proliferation, invasion and migration of colorectal cancer cells

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2021.12437

    Overexpression of HOXC10 partially reverses the inhibitory effect of MTFR2 knockdown on colorectal cancer cell proliferation and migration. (A) Proliferation, (B) clone formation, (C) migration and (D) invasion of HCT116 cells co-transfected with sh-MTFR2-1 and Oe-HOXC10 were in turn detected by the Cell Counting Kit-8, clone formation, wound healing and Transwell assays (magnification, ×100). Relative migration and invasion rates were expressed normalized to the control group. (E) Protein expression levels of invasion- and migration-related proteins in HCT116 cells co-transfected with sh-MTFR2-1 and Oe-HOXC10 were detected by western blotting. **P
    Figure Legend Snippet: Overexpression of HOXC10 partially reverses the inhibitory effect of MTFR2 knockdown on colorectal cancer cell proliferation and migration. (A) Proliferation, (B) clone formation, (C) migration and (D) invasion of HCT116 cells co-transfected with sh-MTFR2-1 and Oe-HOXC10 were in turn detected by the Cell Counting Kit-8, clone formation, wound healing and Transwell assays (magnification, ×100). Relative migration and invasion rates were expressed normalized to the control group. (E) Protein expression levels of invasion- and migration-related proteins in HCT116 cells co-transfected with sh-MTFR2-1 and Oe-HOXC10 were detected by western blotting. **P

    Techniques Used: Over Expression, Migration, Transfection, Cell Counting, Expressing, Western Blot

    MTFR2 knockdown inhibits colorectal cancer cell invasion and migration. (A) Migration and (B) invasion of HCT116 cells transfected with sh-MTFR2-1 were analyzed by wound healing and Transwell assays, respectively (magnification, ×100). Relative migration and invasion rates were expressed normalized to the control group. (C) Expression of invasion- and migration-related proteins in HCT116 cells transfected with sh-MTFR2-1 were detected via western blotting. ***P
    Figure Legend Snippet: MTFR2 knockdown inhibits colorectal cancer cell invasion and migration. (A) Migration and (B) invasion of HCT116 cells transfected with sh-MTFR2-1 were analyzed by wound healing and Transwell assays, respectively (magnification, ×100). Relative migration and invasion rates were expressed normalized to the control group. (C) Expression of invasion- and migration-related proteins in HCT116 cells transfected with sh-MTFR2-1 were detected via western blotting. ***P

    Techniques Used: Migration, Transfection, Expressing, Western Blot

    HOXC10 is upregulated in colorectal cancer cells, and binds to the MTFR2 promoter to activate MTFR2 expression. (A) mRNA and (B) protein expression levels of HOXC10 in HCT116 and HIEC-6 cell lines were analyzed by RT-qPCR and western blotting, respectively. ***P
    Figure Legend Snippet: HOXC10 is upregulated in colorectal cancer cells, and binds to the MTFR2 promoter to activate MTFR2 expression. (A) mRNA and (B) protein expression levels of HOXC10 in HCT116 and HIEC-6 cell lines were analyzed by RT-qPCR and western blotting, respectively. ***P

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot

    16) Product Images from "Anti-helminth compound niclosamide downregulates Wnt Signaling and elicits antitumor responses in tumors with activating APC mutations"

    Article Title: Anti-helminth compound niclosamide downregulates Wnt Signaling and elicits antitumor responses in tumors with activating APC mutations

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-10-3978

    Niclosamide inhibits proliferation of colorectal cancer cell lines and explants
    Figure Legend Snippet: Niclosamide inhibits proliferation of colorectal cancer cell lines and explants

    Techniques Used:

    Niclosamide inhibits Wnt/β-catenin signaling and downregulates dishevelled 2 and β-catenin expression by colorectal cancer cells in vitro
    Figure Legend Snippet: Niclosamide inhibits Wnt/β-catenin signaling and downregulates dishevelled 2 and β-catenin expression by colorectal cancer cells in vitro

    Techniques Used: Expressing, In Vitro

    Combination effect of niclosamide and oxaliplatin on colorectal cancer cell lines and explants
    Figure Legend Snippet: Combination effect of niclosamide and oxaliplatin on colorectal cancer cell lines and explants

    Techniques Used:

    Niclosamide inhibits the growth of colorectal cancers in NOD/SCID mice and downregulates dishevelled 2 and β-catenin expression
    Figure Legend Snippet: Niclosamide inhibits the growth of colorectal cancers in NOD/SCID mice and downregulates dishevelled 2 and β-catenin expression

    Techniques Used: Mouse Assay, Expressing

    Niclosamide inhibits the proliferation of colorectal cancer cell Lines and has minimal toxicity on normal fibroblasts, PBMCs, or immortalized mammary epithelial cells
    Figure Legend Snippet: Niclosamide inhibits the proliferation of colorectal cancer cell Lines and has minimal toxicity on normal fibroblasts, PBMCs, or immortalized mammary epithelial cells

    Techniques Used:

    17) Product Images from "Differential gene expression profile reveals deregulation of pregnancy specific ?1 glycoprotein 9 early during colorectal carcinogenesis"

    Article Title: Differential gene expression profile reveals deregulation of pregnancy specific ?1 glycoprotein 9 early during colorectal carcinogenesis

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-5-66

    PSG9 is exclusively expressed by placental cells . Quantitative RT-PCR analysis of PSG9 transcript (a) and northern blot analysis of different normal tissues (1 μg mRNA/lane) probed with the labelled PSG9 cDNA (b) shows specific expression of PSG9 in placenta. (a-b). Colorectal cancer cell lines were also examined for PSG9 expression level by quantitative- (c) and semi-quantitative RT-PCR (d). Highest expression level of PSG9 was detected in SW480 cells and was lowest in the RKO cell line (c-d).
    Figure Legend Snippet: PSG9 is exclusively expressed by placental cells . Quantitative RT-PCR analysis of PSG9 transcript (a) and northern blot analysis of different normal tissues (1 μg mRNA/lane) probed with the labelled PSG9 cDNA (b) shows specific expression of PSG9 in placenta. (a-b). Colorectal cancer cell lines were also examined for PSG9 expression level by quantitative- (c) and semi-quantitative RT-PCR (d). Highest expression level of PSG9 was detected in SW480 cells and was lowest in the RKO cell line (c-d).

    Techniques Used: Quantitative RT-PCR, Northern Blot, Expressing

    PSG9 expression analysis in Wnt stimulated RKO cells . To determine whether induction of Wnt signaling in cells with wild type APC could induce PSG9 expression, RKO cells with wild type APC and β-catenin were stimulated with either Wnt3a or Kenpaullone (Kenp). After 23 hrs treatment RNA and protein were extracted and processed for PSG9 transcripts expression level by semi-quantitative RT-PCR (a) and β-catenin accumulation by western blot analysis (b). Neither of the treatments nor the RKO-β-catenin S37A (βcat-S37A) stable cell line which expressed constitutively active β-catenin in RKO cells caused expression of PSG9 (a). The SW480 colorectal cancer cell line was used as a positive control (a-b). The RKO cells responded to Wnt stimulation, since cells treated with Wnt3a showed 4-fold induction in luciferase activity compared to untreated cells (c). Axin2, a known downstream target of Wnt signaling, showed 2.4 fold up-regulation in expression as determined by quantitative PCR (d). No PSG9 transcript up-regulation was detected in these cells under these conditions (e). Each sample was analyzed in triplicate.
    Figure Legend Snippet: PSG9 expression analysis in Wnt stimulated RKO cells . To determine whether induction of Wnt signaling in cells with wild type APC could induce PSG9 expression, RKO cells with wild type APC and β-catenin were stimulated with either Wnt3a or Kenpaullone (Kenp). After 23 hrs treatment RNA and protein were extracted and processed for PSG9 transcripts expression level by semi-quantitative RT-PCR (a) and β-catenin accumulation by western blot analysis (b). Neither of the treatments nor the RKO-β-catenin S37A (βcat-S37A) stable cell line which expressed constitutively active β-catenin in RKO cells caused expression of PSG9 (a). The SW480 colorectal cancer cell line was used as a positive control (a-b). The RKO cells responded to Wnt stimulation, since cells treated with Wnt3a showed 4-fold induction in luciferase activity compared to untreated cells (c). Axin2, a known downstream target of Wnt signaling, showed 2.4 fold up-regulation in expression as determined by quantitative PCR (d). No PSG9 transcript up-regulation was detected in these cells under these conditions (e). Each sample was analyzed in triplicate.

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Stable Transfection, Positive Control, Luciferase, Activity Assay, Real-time Polymerase Chain Reaction

    PSG9 is ectopically expressed by cancer cells . A multiple-tumour Northern blot (1 0 μg RNA/lane) revealed over-expression of PSG9 in colon, rectal and uterus cancer. At least three different transcripts were observed (a). Expression of PSG9
    Figure Legend Snippet: PSG9 is ectopically expressed by cancer cells . A multiple-tumour Northern blot (1 0 μg RNA/lane) revealed over-expression of PSG9 in colon, rectal and uterus cancer. At least three different transcripts were observed (a). Expression of PSG9 "isoform a" was examined in a panel of colorectal cancer cases. Forty-nine percent (15/27) of cases showed up-regulation of PSG9 (isoform a) in the tumours compared to corresponding normal tissue (b). However, the expression levels were different between different cases (T; tumour, A; adenoma).

    Techniques Used: Northern Blot, Over Expression, Expressing

    RNA in situ hybridization and IHC analysis of colorectal cancer cases . Sections from sporadic/familial colorectal cancer and placenta (as positive control) were hybridised with Dig-labelled PSG9 RNA probes. Both PSG2 (b) and PSG9 (c) were expressed at a high level in placental tissue. Sense-probes were used as a negative control on placental tissue (a). In microscopic normal epithelial cells from FAP cases, PSG9 expression was detected at the top of crypt (d) (see discussion). PSG9 transcripts (shown as dark blue) were detected at very low levels in normal mucosa (e), adenomas (f), while high expression was detected in tumour cells from the same FAP case (g). In contrast to sporadic cases, PSG9 was detected in normal appearing mucosa in some FAP cases with APC germline mutations, suggesting that dose and level of APC have an impact on PSG9 levels in cells (e, i). A high level of PSG9 was detected in a sporadic case (k), while corresponding normal tissue was negative (i). Tumours and corresponding normal tissue were also examined for β-catenin stabilization by immunostaining (h, j, l). As expected, high levels of β-catenin were detected in all sporadic colorectal tumours (l), while the protein level was less intense in FAP cases (h) where PSG9 up-regulation could be measured (g).
    Figure Legend Snippet: RNA in situ hybridization and IHC analysis of colorectal cancer cases . Sections from sporadic/familial colorectal cancer and placenta (as positive control) were hybridised with Dig-labelled PSG9 RNA probes. Both PSG2 (b) and PSG9 (c) were expressed at a high level in placental tissue. Sense-probes were used as a negative control on placental tissue (a). In microscopic normal epithelial cells from FAP cases, PSG9 expression was detected at the top of crypt (d) (see discussion). PSG9 transcripts (shown as dark blue) were detected at very low levels in normal mucosa (e), adenomas (f), while high expression was detected in tumour cells from the same FAP case (g). In contrast to sporadic cases, PSG9 was detected in normal appearing mucosa in some FAP cases with APC germline mutations, suggesting that dose and level of APC have an impact on PSG9 levels in cells (e, i). A high level of PSG9 was detected in a sporadic case (k), while corresponding normal tissue was negative (i). Tumours and corresponding normal tissue were also examined for β-catenin stabilization by immunostaining (h, j, l). As expected, high levels of β-catenin were detected in all sporadic colorectal tumours (l), while the protein level was less intense in FAP cases (h) where PSG9 up-regulation could be measured (g).

    Techniques Used: RNA In Situ Hybridization, Immunohistochemistry, Positive Control, Negative Control, Expressing, Immunostaining

    18) Product Images from "A phenotypic screen identifies microtubule plus end assembly regulators that can function in mitotic spindle orientation"

    Article Title: A phenotypic screen identifies microtubule plus end assembly regulators that can function in mitotic spindle orientation

    Journal: Cell Cycle

    doi: 10.1080/15384101.2014.1000693

    Chromosomally instable cancer cells with increased microtubule plus end assembly rates exhibit asymmetric monopolar mitotic spindles after Eg5/Kif11 inhibition. ( A ) Example images of symmetric and asymmetric monopolar spindles after treatment with Eg5/Kif11 inhibitors (spindles, anti-α-tubulin: green; kinetochores, CREST: red; chromosomes, DAPI: blue; scale bar, 10 μm). ( B ) Detection and quantification of asymmetric monopolar spindles in chromosomally stable and instable colorectal cancer cell lines after Eg5/Kif11 inhibition. Cells were treated with 2 μM of dimethylenastron (DME) for 3 hrs and monopolar spindles and mitotic chromosome alignment were detected by immunofluorescence microscopy analysis as indicated and quantified (scale bar, 10 μm; mean ± s.d.; 1500–2000 monopolar spindles from 3 independent experiments were evaluated). ( C ) Detection and quantification of asymmetric monopolar spindles in HCT116 cells after loss of CHK2 or upon overexpression of AURKA. Cells were transiently transfected with siRNAs or with plasmids and monopolar spindle formation was evaluated after treatment with DME as in (B). Scale bar, 10 μm. The graphs show mean values ± s.e.m. and 1500–2000 monopolar spindles from 3 independent experiments were evaluated. ( D ) Measurements of mitotic microtubule plus end assembly rates in chromosomally stable HCT116 and RKO cells after treatment with 0.125 nM ​nocodazole. Scatter dot plots show average assembly rates (20 microtubules per cell, mean ± s.e.m., t -test, n = 10–24 cells from 3 independent experiments). ( E ) Quantification of the proportion of HCT116 or RKO cells exhibiting asymmetrically monopolar spindles after treatment with 2 μM DME and 2.5 nM Nocodazole for 3h (mean values ± s.d.; HCT116: 3 independent experiments with a total of 1400 monopolar spindles; RKO: 2 independent experiments with a total of 500 monopolar spindles).
    Figure Legend Snippet: Chromosomally instable cancer cells with increased microtubule plus end assembly rates exhibit asymmetric monopolar mitotic spindles after Eg5/Kif11 inhibition. ( A ) Example images of symmetric and asymmetric monopolar spindles after treatment with Eg5/Kif11 inhibitors (spindles, anti-α-tubulin: green; kinetochores, CREST: red; chromosomes, DAPI: blue; scale bar, 10 μm). ( B ) Detection and quantification of asymmetric monopolar spindles in chromosomally stable and instable colorectal cancer cell lines after Eg5/Kif11 inhibition. Cells were treated with 2 μM of dimethylenastron (DME) for 3 hrs and monopolar spindles and mitotic chromosome alignment were detected by immunofluorescence microscopy analysis as indicated and quantified (scale bar, 10 μm; mean ± s.d.; 1500–2000 monopolar spindles from 3 independent experiments were evaluated). ( C ) Detection and quantification of asymmetric monopolar spindles in HCT116 cells after loss of CHK2 or upon overexpression of AURKA. Cells were transiently transfected with siRNAs or with plasmids and monopolar spindle formation was evaluated after treatment with DME as in (B). Scale bar, 10 μm. The graphs show mean values ± s.e.m. and 1500–2000 monopolar spindles from 3 independent experiments were evaluated. ( D ) Measurements of mitotic microtubule plus end assembly rates in chromosomally stable HCT116 and RKO cells after treatment with 0.125 nM ​nocodazole. Scatter dot plots show average assembly rates (20 microtubules per cell, mean ± s.e.m., t -test, n = 10–24 cells from 3 independent experiments). ( E ) Quantification of the proportion of HCT116 or RKO cells exhibiting asymmetrically monopolar spindles after treatment with 2 μM DME and 2.5 nM Nocodazole for 3h (mean values ± s.d.; HCT116: 3 independent experiments with a total of 1400 monopolar spindles; RKO: 2 independent experiments with a total of 500 monopolar spindles).

    Techniques Used: Inhibition, Immunofluorescence, Microscopy, Over Expression, Transfection

    19) Product Images from "FOXQ1 promotes cancer metastasis by PI3K/AKT signaling regulation in colorectal carcinoma"

    Article Title: FOXQ1 promotes cancer metastasis by PI3K/AKT signaling regulation in colorectal carcinoma

    Journal: American Journal of Translational Research

    doi:

    FOXQ1 expression is up-regulated in human colorectal cancer. A. qRT-PCR analysis of FOXQ1 mRNA levels in various colorectal cancer cell lines. Histograms reporting the levels of FOXQ1 mRNA (normalized to the GAPDH mRNA) as assessed in five representative
    Figure Legend Snippet: FOXQ1 expression is up-regulated in human colorectal cancer. A. qRT-PCR analysis of FOXQ1 mRNA levels in various colorectal cancer cell lines. Histograms reporting the levels of FOXQ1 mRNA (normalized to the GAPDH mRNA) as assessed in five representative

    Techniques Used: Expressing, Quantitative RT-PCR

    Down-expression of FOXQ1 significantly inhibits colorectal cancer cells growth in vivo. A. Control siRNA or siRNA against FOXQ1 were transfected into HCT116 cells and then cells was subcutaneously injected into back flank of nude mice (n = 6 each). Representative
    Figure Legend Snippet: Down-expression of FOXQ1 significantly inhibits colorectal cancer cells growth in vivo. A. Control siRNA or siRNA against FOXQ1 were transfected into HCT116 cells and then cells was subcutaneously injected into back flank of nude mice (n = 6 each). Representative

    Techniques Used: Expressing, In Vivo, Transfection, Injection, Mouse Assay

    Up-regulation of FOXQ1 accelerates cellular migration and invasion in colorectal cancer cells. A. Based on the scratch migration assay, the migration ability of HCT116 and LOVO cells transfected with FOXQ1 was augmented. B. The invasion ability of HCT116
    Figure Legend Snippet: Up-regulation of FOXQ1 accelerates cellular migration and invasion in colorectal cancer cells. A. Based on the scratch migration assay, the migration ability of HCT116 and LOVO cells transfected with FOXQ1 was augmented. B. The invasion ability of HCT116

    Techniques Used: Migration, Transfection

    Knock-down FOXQ1 hinders cellular migration and invasion in colorectal cancer cells. A. LOVO (left panel) and HCT116 (right panel) cells were transfected with either control siRNA or FOXQ1-targeted siRNAs for 48 h prior to wound healing assays. Bars show
    Figure Legend Snippet: Knock-down FOXQ1 hinders cellular migration and invasion in colorectal cancer cells. A. LOVO (left panel) and HCT116 (right panel) cells were transfected with either control siRNA or FOXQ1-targeted siRNAs for 48 h prior to wound healing assays. Bars show

    Techniques Used: Migration, Transfection

    FOXQ1 regulates cell proliferation in colorectal cancer cell lines. A. Western blot shown the protein expression levels of FOXQ1 in the human colorectal cancer cell lines HCT116 and LOVO after transfected with FOXQ1 siRNA or FOXQ1. GAPDH was used as a
    Figure Legend Snippet: FOXQ1 regulates cell proliferation in colorectal cancer cell lines. A. Western blot shown the protein expression levels of FOXQ1 in the human colorectal cancer cell lines HCT116 and LOVO after transfected with FOXQ1 siRNA or FOXQ1. GAPDH was used as a

    Techniques Used: Western Blot, Expressing, Transfection

    20) Product Images from "Glutaminase 1 expression in colorectal cancer cells is induced by hypoxia and required for tumor growth, invasion, and metastatic colonization"

    Article Title: Glutaminase 1 expression in colorectal cancer cells is induced by hypoxia and required for tumor growth, invasion, and metastatic colonization

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-018-1291-5

    Glutaminase 1 (GLS1) deficiency suppresses colorectal cancer cell migration of invasion. a The protein levels of GLS1 in HT29 subclones transfected with lentiviral vectors encoding shRNA targeting GLS1 (shGLS1-1, shGLS1-2, shGLS1-3) and non-targeting control (NTC). b The protein levels of hypoxia-inducible factor (HIF)-1α and GLS1 in HT29 transfected with lentiviral vectors encoding short hairpin RNA (shRNA) targeting both HIF-1α and GLS1 (sh1α/GLS1) that exposed at 20 and 1% O 2 for 48 h. c A total of 5 × 10 5 cells were seeded on top of Matrigel-coated chamber inserts and incubated in serum-free Dulbecco’s modified Eagle’s medium (DMEM) at either 20 or 1% O 2 for 48 h. The number of cells that invaded through the Matrigel to the underside of the filter was determined by staining with crystal violet and counting under bright field microscopy in 10 randomly selected fields. Scale bar = 100 μm. Each group was performed in triplicate, and all the results were repeated by three independent experiments. d Scratch assay was performed to analyze the migration of HT29 subclones (NTC, shHIF-1α, shGLS1, and sh1α/GLS1) by exposure to 20 or 1% O 2 for 72 h. Percentage of cell-free area at indicated time points (0 h, 12 h, 24 h, 48 h, and 72 h) compared with that at 0 h was determined. Each condition was performed in triplicate. All data were repeated by three independent experiments and expressed as mean ± SEM ( n = 3, *** p
    Figure Legend Snippet: Glutaminase 1 (GLS1) deficiency suppresses colorectal cancer cell migration of invasion. a The protein levels of GLS1 in HT29 subclones transfected with lentiviral vectors encoding shRNA targeting GLS1 (shGLS1-1, shGLS1-2, shGLS1-3) and non-targeting control (NTC). b The protein levels of hypoxia-inducible factor (HIF)-1α and GLS1 in HT29 transfected with lentiviral vectors encoding short hairpin RNA (shRNA) targeting both HIF-1α and GLS1 (sh1α/GLS1) that exposed at 20 and 1% O 2 for 48 h. c A total of 5 × 10 5 cells were seeded on top of Matrigel-coated chamber inserts and incubated in serum-free Dulbecco’s modified Eagle’s medium (DMEM) at either 20 or 1% O 2 for 48 h. The number of cells that invaded through the Matrigel to the underside of the filter was determined by staining with crystal violet and counting under bright field microscopy in 10 randomly selected fields. Scale bar = 100 μm. Each group was performed in triplicate, and all the results were repeated by three independent experiments. d Scratch assay was performed to analyze the migration of HT29 subclones (NTC, shHIF-1α, shGLS1, and sh1α/GLS1) by exposure to 20 or 1% O 2 for 72 h. Percentage of cell-free area at indicated time points (0 h, 12 h, 24 h, 48 h, and 72 h) compared with that at 0 h was determined. Each condition was performed in triplicate. All data were repeated by three independent experiments and expressed as mean ± SEM ( n = 3, *** p

    Techniques Used: Migration, Transfection, shRNA, Incubation, Modification, Staining, Microscopy, Wound Healing Assay

    Glutaminase 1 (GLS1) deficiency suppresses colorectal tumor growth and metastatic colonization. a HT29 subclones (2 × 10 6 cells) were implanted into the groin of 6–8-week-old male SCID mice. Primary tumor volume was determined twice per week. After 51 days, the primary tumor was harvested and weighed. *** p
    Figure Legend Snippet: Glutaminase 1 (GLS1) deficiency suppresses colorectal tumor growth and metastatic colonization. a HT29 subclones (2 × 10 6 cells) were implanted into the groin of 6–8-week-old male SCID mice. Primary tumor volume was determined twice per week. After 51 days, the primary tumor was harvested and weighed. *** p

    Techniques Used: Mouse Assay

    Glutaminase 1 (GLS1) high expression is associated with poor prognosis in human cancers. Relative levels of GLS1 mRNA from microarray analysis (normalized log2 ratios) of primary tumor samples relative to adjacent normal tissue from cancer patients (the Cancer Genome Atlas (TCGA) database) are shown. a Light blue, samples from normal colorectal tissue ( n = 50); Dark blue, samples from colorectal adenocarcinomas ( n = 383), p
    Figure Legend Snippet: Glutaminase 1 (GLS1) high expression is associated with poor prognosis in human cancers. Relative levels of GLS1 mRNA from microarray analysis (normalized log2 ratios) of primary tumor samples relative to adjacent normal tissue from cancer patients (the Cancer Genome Atlas (TCGA) database) are shown. a Light blue, samples from normal colorectal tissue ( n = 50); Dark blue, samples from colorectal adenocarcinomas ( n = 383), p

    Techniques Used: Expressing, Microarray

    Immunohistochemical staining of glutaminase 1 (GLS1) in human colorectal carcinoma and adjacent normal colorectal tissue. a , b Representative images of GLS1 ( a ) and GLS2 ( b ) expression in tissue microarrays containing 39 cases of colorectal adenocarcinoma and 15 cases of normal colorectal tissue are shown. The bottom panel shows higher magnification of GLS1 and GLS2 staining (brown). c – e Representative images of GLS1 in tissue microarrays containing tumor samples from 85 colorectal cancer patients (24 patients with the presence of lymph node metastasis) are shown, respectively. c Adjacent normal colorectal tissue; d Primary colorectal tumor tissue; e Lymph node metastasis. The bottom panel shows higher magnification of GLS1 staining. Scale bar = 200 μm; f Image analysis was performed to determine the percentage of GLS1-positive area per field under × 40 magnification based on six random fields per section ( n = 24). For bar graph, one-way analysis of variance (ANOVA; mean ± SEM) was used to determine the p- value, *** p
    Figure Legend Snippet: Immunohistochemical staining of glutaminase 1 (GLS1) in human colorectal carcinoma and adjacent normal colorectal tissue. a , b Representative images of GLS1 ( a ) and GLS2 ( b ) expression in tissue microarrays containing 39 cases of colorectal adenocarcinoma and 15 cases of normal colorectal tissue are shown. The bottom panel shows higher magnification of GLS1 and GLS2 staining (brown). c – e Representative images of GLS1 in tissue microarrays containing tumor samples from 85 colorectal cancer patients (24 patients with the presence of lymph node metastasis) are shown, respectively. c Adjacent normal colorectal tissue; d Primary colorectal tumor tissue; e Lymph node metastasis. The bottom panel shows higher magnification of GLS1 staining. Scale bar = 200 μm; f Image analysis was performed to determine the percentage of GLS1-positive area per field under × 40 magnification based on six random fields per section ( n = 24). For bar graph, one-way analysis of variance (ANOVA; mean ± SEM) was used to determine the p- value, *** p

    Techniques Used: Immunohistochemistry, Staining, Expressing

    Hypoxia induces glutaminase 1 (GLS1) expression in a HIF-1α-dependent manner in colorectal cancer cell lines. a GLS1 protein expression in colorectal cancer cell lines. b Reverse transcription and quantitative real-time PCR (RT-qPCR) was performed to quantify GLS1 mRNA levels in colorectal cancer cell lines (HT29 and Caco2) following exposure to 20 or 1% O 2 for 24 h. For each sample, the expression of GLS1 mRNA was quantified relative to 18S rRNA and then normalized to the result obtained from cells at 20% O 2 . Statistical analysis was performed before normalization. Data are shown as mean ± SEM; n = 3. ** p
    Figure Legend Snippet: Hypoxia induces glutaminase 1 (GLS1) expression in a HIF-1α-dependent manner in colorectal cancer cell lines. a GLS1 protein expression in colorectal cancer cell lines. b Reverse transcription and quantitative real-time PCR (RT-qPCR) was performed to quantify GLS1 mRNA levels in colorectal cancer cell lines (HT29 and Caco2) following exposure to 20 or 1% O 2 for 24 h. For each sample, the expression of GLS1 mRNA was quantified relative to 18S rRNA and then normalized to the result obtained from cells at 20% O 2 . Statistical analysis was performed before normalization. Data are shown as mean ± SEM; n = 3. ** p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

    21) Product Images from "Oncolytic Activity of a Recombinant Measles Virus, Blind to Signaling Lymphocyte Activation Molecule, Against Colorectal Cancer Cells"

    Article Title: Oncolytic Activity of a Recombinant Measles Virus, Blind to Signaling Lymphocyte Activation Molecule, Against Colorectal Cancer Cells

    Journal: Scientific Reports

    doi: 10.1038/srep24572

    Infectivity and killing activity of rMV-EGFP-SLAMblind in colorectal cancer cells. ( a ) Representative fluorescence microscopy data. Cells were inoculated with rMV-EGFP-SLAMblind at MOI 2 and incubated for 72 h. Fluorescence microscopy was used to detect infection with rMV-EGFP-SLAMblind. Shown are representative data of three independent experiments. Original magnification, 20× objective lens. Bar, 100 μm. Changes in cell viability in nectin-4-positive ( b ) and nectin-4-negative ( c ) colorectal cancer cells. Cells were inoculated with rMV-EGFP-SLAMblind at MOI 2, and a WST assay was conducted on the indicated dpi. Each datum represents the mean ± SD of three independent experiments.
    Figure Legend Snippet: Infectivity and killing activity of rMV-EGFP-SLAMblind in colorectal cancer cells. ( a ) Representative fluorescence microscopy data. Cells were inoculated with rMV-EGFP-SLAMblind at MOI 2 and incubated for 72 h. Fluorescence microscopy was used to detect infection with rMV-EGFP-SLAMblind. Shown are representative data of three independent experiments. Original magnification, 20× objective lens. Bar, 100 μm. Changes in cell viability in nectin-4-positive ( b ) and nectin-4-negative ( c ) colorectal cancer cells. Cells were inoculated with rMV-EGFP-SLAMblind at MOI 2, and a WST assay was conducted on the indicated dpi. Each datum represents the mean ± SD of three independent experiments.

    Techniques Used: Infection, Activity Assay, Fluorescence, Microscopy, Incubation, WST Assay

    Expression of MV receptors on colorectal cancer cells. ( a ) Flow-cytometric analysis of cell-surface proteins that are associated with MV entry. A total number of 10 6 cells were stained with each primary and secondary Ab (black line). For nectin-4 detection, mAb (clone N4.61) was used in this experiment. The grey histogram indicates the IgG control for each cell line. Shown are representative data of three independent experiments. ( b ) RT-PCR analysis of each colorectal cancer cell line. Representative data of the electrophoresis (left) as well as relative expression level of nectin-4 based on the qRT-PCR analysis (right) is presented. In the qRT-PCR analysis, expression level of nectin-4 was normalized to GAPDH , and the average ± SD of three independent experiments is indicated.
    Figure Legend Snippet: Expression of MV receptors on colorectal cancer cells. ( a ) Flow-cytometric analysis of cell-surface proteins that are associated with MV entry. A total number of 10 6 cells were stained with each primary and secondary Ab (black line). For nectin-4 detection, mAb (clone N4.61) was used in this experiment. The grey histogram indicates the IgG control for each cell line. Shown are representative data of three independent experiments. ( b ) RT-PCR analysis of each colorectal cancer cell line. Representative data of the electrophoresis (left) as well as relative expression level of nectin-4 based on the qRT-PCR analysis (right) is presented. In the qRT-PCR analysis, expression level of nectin-4 was normalized to GAPDH , and the average ± SD of three independent experiments is indicated.

    Techniques Used: Expressing, Flow Cytometry, Staining, Reverse Transcription Polymerase Chain Reaction, Electrophoresis, Quantitative RT-PCR

    22) Product Images from "MicroRNA-513a-3p regulates colorectal cancer cell metabolism via targeting hexokinase 2"

    Article Title: MicroRNA-513a-3p regulates colorectal cancer cell metabolism via targeting hexokinase 2

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2020.8727

    MiR-513a-3p inhibited colorectal cancer cell proliferation. (A) Transfection of miR-513a-3p mimic elevated miR-513a-3p expression in HCT116 cells. (B) Transfection of miR-513a-3p mimic elevated miR-513a-3p expression in SW480 cells. (C) Overexpression of miR-513a-3p reduced cell proliferation of HCT116 cells. (D) Overexpression of miR-513a-3p reduced cell proliferation of SW480 cells. *** P
    Figure Legend Snippet: MiR-513a-3p inhibited colorectal cancer cell proliferation. (A) Transfection of miR-513a-3p mimic elevated miR-513a-3p expression in HCT116 cells. (B) Transfection of miR-513a-3p mimic elevated miR-513a-3p expression in SW480 cells. (C) Overexpression of miR-513a-3p reduced cell proliferation of HCT116 cells. (D) Overexpression of miR-513a-3p reduced cell proliferation of SW480 cells. *** P

    Techniques Used: Transfection, Expressing, Over Expression

    MiR-513a-3p inhibited glycolysis of colorectal cancer cells. (A) Overexpression of miR-513a-3p inhibited glucose uptake of HCT116 cells. (B) Overexpression of miR-513a-3p inhibited glucose uptake of SW480 cells. (C) Overexpression of miR-513a-3p decreased lactate levels in culture medium of HCT116 cells. (D) Overexpression of miR-513a-3p decreased lactate levels in culture medium of SW480 cells. ** P
    Figure Legend Snippet: MiR-513a-3p inhibited glycolysis of colorectal cancer cells. (A) Overexpression of miR-513a-3p inhibited glucose uptake of HCT116 cells. (B) Overexpression of miR-513a-3p inhibited glucose uptake of SW480 cells. (C) Overexpression of miR-513a-3p decreased lactate levels in culture medium of HCT116 cells. (D) Overexpression of miR-513a-3p decreased lactate levels in culture medium of SW480 cells. ** P

    Techniques Used: Over Expression

    MiR-513a-3p decreased HK2 expression in colorectal cancer cells. (A) Among a panel of glycolysis associated genes, miR-513a-3p overexpression decreased HK2 mRNA levels in HCT116 cells. (B) Among a panel of glycolysis associated genes, miR-513a-3p overexpression decreased HK2 mRNA levels in SW480 cells. (C) Overexpression of miR-513a-3p decreased HK2 protein expression in HCT116 and SW480 cells. (D) Quantitative analysis of HK2 protein expression in C. ** P
    Figure Legend Snippet: MiR-513a-3p decreased HK2 expression in colorectal cancer cells. (A) Among a panel of glycolysis associated genes, miR-513a-3p overexpression decreased HK2 mRNA levels in HCT116 cells. (B) Among a panel of glycolysis associated genes, miR-513a-3p overexpression decreased HK2 mRNA levels in SW480 cells. (C) Overexpression of miR-513a-3p decreased HK2 protein expression in HCT116 and SW480 cells. (D) Quantitative analysis of HK2 protein expression in C. ** P

    Techniques Used: Expressing, Over Expression

    HK2 was pivotal for regulation of cell proliferation and glycolysis by miR-513a-3p in colorectal cancer cells. (A) Western blotting showed that transfection of pcDNA3-HK2 elevated HK2 protein expression in HCT116 and SW480 cells. (B) Quantitative analysis of HK2 expression in A. (C and D) The cell proliferation assay showed that the overexpression of HK2 reversed cell proliferation inhibition led by miR-513a-3p mimic in HCT116 and SW480 cells. (E and F) Overexpression of HK2 reversed reduction of glucose uptake induced by miR-513a-3p mimic in HCT116 and SW480 cells. (G and H) Overexpression of HK2 reversed reduction of lactate levels induced by miR-513a-3p mimic in culture medium of HCT116 and SW480 cells. * P
    Figure Legend Snippet: HK2 was pivotal for regulation of cell proliferation and glycolysis by miR-513a-3p in colorectal cancer cells. (A) Western blotting showed that transfection of pcDNA3-HK2 elevated HK2 protein expression in HCT116 and SW480 cells. (B) Quantitative analysis of HK2 expression in A. (C and D) The cell proliferation assay showed that the overexpression of HK2 reversed cell proliferation inhibition led by miR-513a-3p mimic in HCT116 and SW480 cells. (E and F) Overexpression of HK2 reversed reduction of glucose uptake induced by miR-513a-3p mimic in HCT116 and SW480 cells. (G and H) Overexpression of HK2 reversed reduction of lactate levels induced by miR-513a-3p mimic in culture medium of HCT116 and SW480 cells. * P

    Techniques Used: Western Blot, Transfection, Expressing, Proliferation Assay, Over Expression, Inhibition

    HK2 mRNA levels were negatively correlated with miR-513a-3p levels in tumors and normal tissues from colorectal cancer patients. (A) RT-qPCR showed that HK2 mRNA expression was elevated in tumors compared with matched normal tissues from 30 patients with colorectal cancer. (B) The Pearson correlation analysis suggested that miR-513a-3p levels were negatively correlated with HK2 mRNA expression in tumors and normal tissues from 30 patients with colorectal cancer. *** P
    Figure Legend Snippet: HK2 mRNA levels were negatively correlated with miR-513a-3p levels in tumors and normal tissues from colorectal cancer patients. (A) RT-qPCR showed that HK2 mRNA expression was elevated in tumors compared with matched normal tissues from 30 patients with colorectal cancer. (B) The Pearson correlation analysis suggested that miR-513a-3p levels were negatively correlated with HK2 mRNA expression in tumors and normal tissues from 30 patients with colorectal cancer. *** P

    Techniques Used: Quantitative RT-PCR, Expressing

    23) Product Images from "Elevated Tolerance to Aneuploidy in Cancer Cells: Estimating the Fitness Effects of Chromosome Number Alterations by In Silico Modelling of Somatic Genome Evolution"

    Article Title: Elevated Tolerance to Aneuploidy in Cancer Cells: Estimating the Fitness Effects of Chromosome Number Alterations by In Silico Modelling of Somatic Genome Evolution

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070445

    Modelling the distribution of aneuploidy burden in human cancers. (A) Reported cytogenetic data from the Mitelman Database of Chromosome Aberrations and Gene Fusions in Cancer show a log-linear relationship between the relative prevalence and the number of numerical aberrations per tumour (Nnapt), with highly similar distributions for Wilms tumour (WT) and colorectal cancer (CRC). (B) Modelling of a certain number of cancer stemlines arising in the same number of patients. Each stemline is assumed to derive from a diploid cell (having 0 numerical aberrations) and is allowed to proliferate for a maximum of 2000 generations (G), when the overall distribution of numerical aberrations is sampled. Stemlines accumulate numerical aberrations at a certain mis-segregation rate ( p ) and are subject to aneuploidy-dependent selection at a certain degree ( s ), which may in turn result in termination of the stemline (horizontal dumbbell), corresponding to the end of clonal expansion. Because this may result in regression of tumorigenesis at an early stage, cases where stemlines were thus terminated were removed from sampling. (C) Simulated distribution of tumour cases with a certain number of numerical aberrations as the tumour cohort is sampled at generations 1–2000 in a setting where tumours harbour an elevated mis-segregation rate in the absence of negative selection against aneuploid cells (see main text for details). This will result in a binomial-like distribution already after 100 generations, the modal value of which increases with time, in contrast to the actual distribution in human tumours (compare to 6A).
    Figure Legend Snippet: Modelling the distribution of aneuploidy burden in human cancers. (A) Reported cytogenetic data from the Mitelman Database of Chromosome Aberrations and Gene Fusions in Cancer show a log-linear relationship between the relative prevalence and the number of numerical aberrations per tumour (Nnapt), with highly similar distributions for Wilms tumour (WT) and colorectal cancer (CRC). (B) Modelling of a certain number of cancer stemlines arising in the same number of patients. Each stemline is assumed to derive from a diploid cell (having 0 numerical aberrations) and is allowed to proliferate for a maximum of 2000 generations (G), when the overall distribution of numerical aberrations is sampled. Stemlines accumulate numerical aberrations at a certain mis-segregation rate ( p ) and are subject to aneuploidy-dependent selection at a certain degree ( s ), which may in turn result in termination of the stemline (horizontal dumbbell), corresponding to the end of clonal expansion. Because this may result in regression of tumorigenesis at an early stage, cases where stemlines were thus terminated were removed from sampling. (C) Simulated distribution of tumour cases with a certain number of numerical aberrations as the tumour cohort is sampled at generations 1–2000 in a setting where tumours harbour an elevated mis-segregation rate in the absence of negative selection against aneuploid cells (see main text for details). This will result in a binomial-like distribution already after 100 generations, the modal value of which increases with time, in contrast to the actual distribution in human tumours (compare to 6A).

    Techniques Used: Selection, Sampling

    Distribution of numerical aberrations in human cancers predicted by the presence/absence of chromosomal instability and aneuploidy-dependent selection. The expected distributions according to different conditions of chromosomal mis-segregation and selection were predicted by simulations as described in Figure 6 . In each graph, the reported data for Wilms tumour (WT, black circles) and colorectal cancer (CRC, red circles) are included for comparison. (A) The expected distribution (open blue circles) of numerical changes in a setting with chromosomal instability (CIN) in the absence of aneuploidy-dependent negative selection ( s ) is binomial-like with a high modal number of aberrations per tumour. All data points from 10 independent simulations were included. (B) A setting with CIN present but with s values similar to normal fibroblasts, results in a distribution with far fewer aneusomies than reported in WT and CRC. The same conditions, but with absence of CIN (C and D) result in similar distributions as when CIN is included, but with fewer abnormalities. (E) Attenuated aneusomy-dependent negative selection ( s set as a span corresponding to the magnitude found in cancer cell lines) and with CIN present predicts a distribution of numerical changes highly similar to reported data. (F) In contrast, attenuated negative selection combined with absence of CIN results in distribution skewed towards fewer numerical aberrations than observed in WT and CRC.
    Figure Legend Snippet: Distribution of numerical aberrations in human cancers predicted by the presence/absence of chromosomal instability and aneuploidy-dependent selection. The expected distributions according to different conditions of chromosomal mis-segregation and selection were predicted by simulations as described in Figure 6 . In each graph, the reported data for Wilms tumour (WT, black circles) and colorectal cancer (CRC, red circles) are included for comparison. (A) The expected distribution (open blue circles) of numerical changes in a setting with chromosomal instability (CIN) in the absence of aneuploidy-dependent negative selection ( s ) is binomial-like with a high modal number of aberrations per tumour. All data points from 10 independent simulations were included. (B) A setting with CIN present but with s values similar to normal fibroblasts, results in a distribution with far fewer aneusomies than reported in WT and CRC. The same conditions, but with absence of CIN (C and D) result in similar distributions as when CIN is included, but with fewer abnormalities. (E) Attenuated aneusomy-dependent negative selection ( s set as a span corresponding to the magnitude found in cancer cell lines) and with CIN present predicts a distribution of numerical changes highly similar to reported data. (F) In contrast, attenuated negative selection combined with absence of CIN results in distribution skewed towards fewer numerical aberrations than observed in WT and CRC.

    Techniques Used: Selection

    24) Product Images from "Activation of the mTOR Pathway by Oxaliplatin in the Treatment of Colorectal Cancer Liver Metastasis"

    Article Title: Activation of the mTOR Pathway by Oxaliplatin in the Treatment of Colorectal Cancer Liver Metastasis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0169439

    Unsupervised hierarchical clustering on 39 colorectal cancer liver metastasis samples were performed with oxaliplatin treated samples boxed. Unsupervised hierarchical clustering showed that oxaliplatin treatment did not influence the clustering of the 39 samples.
    Figure Legend Snippet: Unsupervised hierarchical clustering on 39 colorectal cancer liver metastasis samples were performed with oxaliplatin treated samples boxed. Unsupervised hierarchical clustering showed that oxaliplatin treatment did not influence the clustering of the 39 samples.

    Techniques Used:

    Activation of the mTOR pathway by oxaliplatin in vivo . The level of phosphorylated p70 S6 kinase was increased in colorectal cancer PDXs by Day 2.
    Figure Legend Snippet: Activation of the mTOR pathway by oxaliplatin in vivo . The level of phosphorylated p70 S6 kinase was increased in colorectal cancer PDXs by Day 2.

    Techniques Used: Activation Assay, In Vivo

    Activation of the mTOR pathway by oxaliplatin in vitro . A. The level of phosphorylated p70 S6 kinase was increased in all five ATCC cell lines (HCT15, DLD-1, LoVo, HCT116, HT29 and Colo 205) by Day 3. B. The level of phosphorylated p70 S6 kinase was increased in all early passage colorectal cancer cell lines (CRC057, CRC119 and CRC240) by Day 3 or 4.
    Figure Legend Snippet: Activation of the mTOR pathway by oxaliplatin in vitro . A. The level of phosphorylated p70 S6 kinase was increased in all five ATCC cell lines (HCT15, DLD-1, LoVo, HCT116, HT29 and Colo 205) by Day 3. B. The level of phosphorylated p70 S6 kinase was increased in all early passage colorectal cancer cell lines (CRC057, CRC119 and CRC240) by Day 3 or 4.

    Techniques Used: Activation Assay, In Vitro

    Oxaliplatin and Rapamycin Synergy Graphs were performed to determine the combination index (CI). A . The growth of ATCC colorectal cell lines HCT15, DLD-1, LoVo, HCT116, HT29 and Colo 205 in the presence of oxaliplatin and rapamycin as single agents and in combination was analyzed by cytotoxicity assays (see materials and methods ) to determine the dual-effect of the two agents on the cell lines. Mean Combination Index (CI) values (from three experiments) at 25%, 50%, 75% and 90% is plotted for each cell line. (CI
    Figure Legend Snippet: Oxaliplatin and Rapamycin Synergy Graphs were performed to determine the combination index (CI). A . The growth of ATCC colorectal cell lines HCT15, DLD-1, LoVo, HCT116, HT29 and Colo 205 in the presence of oxaliplatin and rapamycin as single agents and in combination was analyzed by cytotoxicity assays (see materials and methods ) to determine the dual-effect of the two agents on the cell lines. Mean Combination Index (CI) values (from three experiments) at 25%, 50%, 75% and 90% is plotted for each cell line. (CI

    Techniques Used:

    25) 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 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

    26) Product Images from "m(6)A demethylase ALKBH5 inhibits cell proliferation and the metastasis of colorectal cancer by regulating the FOXO3/miR-21/SPRY2 axis"

    Article Title: m(6)A demethylase ALKBH5 inhibits cell proliferation and the metastasis of colorectal cancer by regulating the FOXO3/miR-21/SPRY2 axis

    Journal: American Journal of Translational Research

    doi:

    The expression levels of ALKBH5 in the clinical tissues and cell lines of colorectal cancer. A: The mRNA levels of ALKBH5 in the tumor tissues and adjacent normal tissues of colorectal cancer (n=36) were determined using RT-qPCR; B: The protein levels of ALKBH5 in the tumor tissues and adjacent normal tissues of colorectal cancer (n=36) were determined using western blot; C: The association between ALKBH5 and the stages of colorectal cancer (n=36); D: The Kaplan Meier plotter was applied to analyze the survival differences between patients with high and low expressions of ALKBH5 (n=36); E: The mRNA levels of ALKBH5 in the cell lines of colorectal cancer (HCT-116, Caco-2, SW480, and SW620) and the normal fetal colonic mucosa cell line (FHC) were determined using RT-qPCR (n=3); F: The protein levels of ALKBH5 in the cell lines of colorectal cancer (HCT-116, Caco-2, SW480, and SW620) and the normal fetal colonic mucosa cell line (FHC) were examined using western blot (n=3). The asterisk represents a significant difference: *P
    Figure Legend Snippet: The expression levels of ALKBH5 in the clinical tissues and cell lines of colorectal cancer. A: The mRNA levels of ALKBH5 in the tumor tissues and adjacent normal tissues of colorectal cancer (n=36) were determined using RT-qPCR; B: The protein levels of ALKBH5 in the tumor tissues and adjacent normal tissues of colorectal cancer (n=36) were determined using western blot; C: The association between ALKBH5 and the stages of colorectal cancer (n=36); D: The Kaplan Meier plotter was applied to analyze the survival differences between patients with high and low expressions of ALKBH5 (n=36); E: The mRNA levels of ALKBH5 in the cell lines of colorectal cancer (HCT-116, Caco-2, SW480, and SW620) and the normal fetal colonic mucosa cell line (FHC) were determined using RT-qPCR (n=3); F: The protein levels of ALKBH5 in the cell lines of colorectal cancer (HCT-116, Caco-2, SW480, and SW620) and the normal fetal colonic mucosa cell line (FHC) were examined using western blot (n=3). The asterisk represents a significant difference: *P

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot

    ALKBH5 suppresses the tumor progression of colorectal cancer. A: The expression of ALKBH5 in HCT-116 and SW480 cells with an overexpression of ALKBH5 was determined using western blot and RT-qPCR (n=3); B: Cell viability was determined using CCK8 assays in HCT-116 and SW480 cells with an overexpression of ALKBH5 (n=3); C: Cell proliferation was determined using colony formation assay in the HCT-116 and SW480 cells with an overexpression of ALKBH5 (scale bar: 50 µM, n=3); D: Cell migration was determined using wound healing assays in the HCT-116 and SW480 cells with overexpressions of ALKBH5 (scale bar: 100 µM, 200×, n=3); E: Cell invasion was determined using transwell assays in the HCT-116 and SW480 cells with an overexpression of ALKBH5 (Scale bar: 100 µM, 200×, n=3). The asterisk represents a significant difference: *P
    Figure Legend Snippet: ALKBH5 suppresses the tumor progression of colorectal cancer. A: The expression of ALKBH5 in HCT-116 and SW480 cells with an overexpression of ALKBH5 was determined using western blot and RT-qPCR (n=3); B: Cell viability was determined using CCK8 assays in HCT-116 and SW480 cells with an overexpression of ALKBH5 (n=3); C: Cell proliferation was determined using colony formation assay in the HCT-116 and SW480 cells with an overexpression of ALKBH5 (scale bar: 50 µM, n=3); D: Cell migration was determined using wound healing assays in the HCT-116 and SW480 cells with overexpressions of ALKBH5 (scale bar: 100 µM, 200×, n=3); E: Cell invasion was determined using transwell assays in the HCT-116 and SW480 cells with an overexpression of ALKBH5 (Scale bar: 100 µM, 200×, n=3). The asterisk represents a significant difference: *P

    Techniques Used: Expressing, Over Expression, Western Blot, Quantitative RT-PCR, Colony Assay, Migration

    27) Product Images from "FSTL1 interacts with VIM and promotes colorectal cancer metastasis via activating the focal adhesion signalling pathway"

    Article Title: FSTL1 interacts with VIM and promotes colorectal cancer metastasis via activating the focal adhesion signalling pathway

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-018-0695-6

    FSTL1 is up-regulated in CRC and correlates with poor prognosis. Expression analyses of FSTL1 protein and mRNA in CRC cell lines by a western blotting and b qPCR. FSTL1 protein levels were normalised to the relative expression of FHC. FSTL1 mRNA expression was quantified by qPCR and normalised to GAPDH. Error bars represent the mean ± S.D. ( n = 3). c Immunohistochemistry (IHC) staining in 130 paraffin-embedded CRC tissues sections. (i) Strong expression (+++) in CRC. (ii) Weak expression (+) in CRC. (iii) Weak expression (+) in adjacent normal tissues. (iv) Scored negative for expression (−) in adjacent normal colorectal tissues. d Comparison of FSTL1 expression scores in CRC tissues (Tumour) with adjacent non-tumour tissues (Normal), P
    Figure Legend Snippet: FSTL1 is up-regulated in CRC and correlates with poor prognosis. Expression analyses of FSTL1 protein and mRNA in CRC cell lines by a western blotting and b qPCR. FSTL1 protein levels were normalised to the relative expression of FHC. FSTL1 mRNA expression was quantified by qPCR and normalised to GAPDH. Error bars represent the mean ± S.D. ( n = 3). c Immunohistochemistry (IHC) staining in 130 paraffin-embedded CRC tissues sections. (i) Strong expression (+++) in CRC. (ii) Weak expression (+) in CRC. (iii) Weak expression (+) in adjacent normal tissues. (iv) Scored negative for expression (−) in adjacent normal colorectal tissues. d Comparison of FSTL1 expression scores in CRC tissues (Tumour) with adjacent non-tumour tissues (Normal), P

    Techniques Used: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Immunohistochemistry, Staining

    28) Product Images from "Modifying Effects of Glucose and Insulin/Insulin-Like Growth Factors on Colon Cancer Cells"

    Article Title: Modifying Effects of Glucose and Insulin/Insulin-Like Growth Factors on Colon Cancer Cells

    Journal: Frontiers in Oncology

    doi: 10.3389/fonc.2021.645732

    mRNA expression levels of the IGF-I (A) , IGF-II (B) , IGF-IR (C) , IGF-IIR (D) , > IR-A (E) and IR-B (F) (expressed as relative mRNA expression, normalized to the house-keeping genes HPRT , GUSB and ACTB ) in four human colorectal cancer cell lines. Two different conditions were tested: medium containing 5 mmol/L glucose (open bars) or 25 mmol/L glucose (black bars). Values represent the mean ± SEM of two independent experiments. *p
    Figure Legend Snippet: mRNA expression levels of the IGF-I (A) , IGF-II (B) , IGF-IR (C) , IGF-IIR (D) , > IR-A (E) and IR-B (F) (expressed as relative mRNA expression, normalized to the house-keeping genes HPRT , GUSB and ACTB ) in four human colorectal cancer cell lines. Two different conditions were tested: medium containing 5 mmol/L glucose (open bars) or 25 mmol/L glucose (black bars). Values represent the mean ± SEM of two independent experiments. *p

    Techniques Used: Expressing

    mRNA expression levels of the IGFBP-1 (A) , IGFBP-2 (B) , IGFBP-3 (C) and IGFBP-6 (D) (expressed as relative mRNA expression, normalized to the house-keeping genes HPRT, GUSB and ACTB ) in four human colorectal cancer cell lines. Two different conditions were tested: medium containing 5 mmol/L glucose (open bars) or 25 mmol/L glucose (black bars). Values represent the mean ± SEM of two independent experiments. *p
    Figure Legend Snippet: mRNA expression levels of the IGFBP-1 (A) , IGFBP-2 (B) , IGFBP-3 (C) and IGFBP-6 (D) (expressed as relative mRNA expression, normalized to the house-keeping genes HPRT, GUSB and ACTB ) in four human colorectal cancer cell lines. Two different conditions were tested: medium containing 5 mmol/L glucose (open bars) or 25 mmol/L glucose (black bars). Values represent the mean ± SEM of two independent experiments. *p

    Techniques Used: Expressing

    29) Product Images from "DACT3 is an epigenetic regulator of Wnt/β-catenin signaling in colorectal cancer and is a therapeutic target of histone modifications"

    Article Title: DACT3 is an epigenetic regulator of Wnt/β-catenin signaling in colorectal cancer and is a therapeutic target of histone modifications

    Journal:

    doi: 10.1016/j.ccr.2008.04.019

    Combination of DZNep and TSA results in blockade of Wnt/β-catenin signaling and massive apoptosis in colorectal cancer cells
    Figure Legend Snippet: Combination of DZNep and TSA results in blockade of Wnt/β-catenin signaling and massive apoptosis in colorectal cancer cells

    Techniques Used:

    Effects of DACT3 overexpression on Dvl2 and β-catenin in colorectal cancer cells
    Figure Legend Snippet: Effects of DACT3 overexpression on Dvl2 and β-catenin in colorectal cancer cells

    Techniques Used: Over Expression

    30) Product Images from "GLUT1 gene is a potential hypoxic marker in colorectal cancer patients"

    Article Title: GLUT1 gene is a potential hypoxic marker in colorectal cancer patients

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-9-241

    Relative lactate concentration curves of two colorectal cancer cell lines . Supernatants from the two colorectal cancer cell lines SW480 and SW620. Control specimens were acquired from the supernatants of culture medium only. The concentrations of lactate were examined after incubation for 24 h and 48 h in hypoxia and normoxia conditions. Relative elevated lactate concentrations of SW480 cell lines were 3.24-fold (hypoxia/normoxia: 41.63/12.84; after 24 h incubation) and 3.36-fold (hypoxia/normoxia: 65.70/19.56; after 48 h incubation); however, there were 3.06-fold (hypoxia/normoxia: 30.78/10.05; after 24 h incubation) and 3.17-fold (hypoxia/normoxia: 70.45/22.23; after 48 h incubation) elevated lactate in SW620 cell lines. Values are means ± SD of 3 independent experiments with triplicate dishes.
    Figure Legend Snippet: Relative lactate concentration curves of two colorectal cancer cell lines . Supernatants from the two colorectal cancer cell lines SW480 and SW620. Control specimens were acquired from the supernatants of culture medium only. The concentrations of lactate were examined after incubation for 24 h and 48 h in hypoxia and normoxia conditions. Relative elevated lactate concentrations of SW480 cell lines were 3.24-fold (hypoxia/normoxia: 41.63/12.84; after 24 h incubation) and 3.36-fold (hypoxia/normoxia: 65.70/19.56; after 48 h incubation); however, there were 3.06-fold (hypoxia/normoxia: 30.78/10.05; after 24 h incubation) and 3.17-fold (hypoxia/normoxia: 70.45/22.23; after 48 h incubation) elevated lactate in SW620 cell lines. Values are means ± SD of 3 independent experiments with triplicate dishes.

    Techniques Used: Concentration Assay, Incubation

    Comparison of the hypoxia- and glycolysis-associated gene expression intensity between human CRC (Case No. 6) and normal tissue . The results showed the relative expression ratio of sixteen genes. The overexpression of GLUT1, HIF-1α, and HIF-2α are presented in these human colorectal cancer tissue samples.
    Figure Legend Snippet: Comparison of the hypoxia- and glycolysis-associated gene expression intensity between human CRC (Case No. 6) and normal tissue . The results showed the relative expression ratio of sixteen genes. The overexpression of GLUT1, HIF-1α, and HIF-2α are presented in these human colorectal cancer tissue samples.

    Techniques Used: Expressing, Over Expression

    Schematic representation of membrane array and comparison of gene expression patterns between a human colorectal cancer patient (Case No. 6) and healthy control . (A) Schematic representation of membrane array with 16 target genes, one housekeeping gene (β-actin), one bacteria gene (TB), and blank control. Sixteen target genes (i.e., GLUT1, HK1, GPI, PGK1, PGK2, ENO2, PKM2, uPA, CA9, CA12, TP, bFGF, COX-2, HIF-1α, HIF-2α, and VEGF). The position of the correlation of the blank and positive control (β-actin) as well as the negative control ( Mycobacterium tuberculosis ; TB) in the nylon membrane are pointed out by spots. The relevant positions are the red area (β-actin), blue area (blank), green area (TB), yellow area (glycolysis-related genes), and pink area (hypoxia-related genes). (B) A triplicate set of 16 molecular markers for colorectal cancer was blotted on the nylon membrane. In addition, a housekeeping gene and a bacterial gene serving as positive and negative controls, respectively, were also blotted on the membrane.
    Figure Legend Snippet: Schematic representation of membrane array and comparison of gene expression patterns between a human colorectal cancer patient (Case No. 6) and healthy control . (A) Schematic representation of membrane array with 16 target genes, one housekeeping gene (β-actin), one bacteria gene (TB), and blank control. Sixteen target genes (i.e., GLUT1, HK1, GPI, PGK1, PGK2, ENO2, PKM2, uPA, CA9, CA12, TP, bFGF, COX-2, HIF-1α, HIF-2α, and VEGF). The position of the correlation of the blank and positive control (β-actin) as well as the negative control ( Mycobacterium tuberculosis ; TB) in the nylon membrane are pointed out by spots. The relevant positions are the red area (β-actin), blue area (blank), green area (TB), yellow area (glycolysis-related genes), and pink area (hypoxia-related genes). (B) A triplicate set of 16 molecular markers for colorectal cancer was blotted on the nylon membrane. In addition, a housekeeping gene and a bacterial gene serving as positive and negative controls, respectively, were also blotted on the membrane.

    Techniques Used: Membrane Array, Expressing, Positive Control, Negative Control

    31) Product Images from "Therapeutic potential of targeting S100A11 in malignant pleural mesothelioma"

    Article Title: Therapeutic potential of targeting S100A11 in malignant pleural mesothelioma

    Journal: Oncogenesis

    doi: 10.1038/s41389-017-0017-3

    High secretion and overexpression of S100A11 in MPM. a Concentration of secreted S100A11 in the culture media of various cell lines, as determined by ELISA. The secretion level of S100A11 in all MPM cell lines, except for 211H, is higher than that of NM. 211H MSTO-211H, NM normal mesothelial cell, LC lung cancer, GC gastric cancer, CRC colorectal cancer, BC breast cancer. b Protein level of S100A11 was higher in MPM cell lines, compared to NM. NM normal mesothelial cell. c Immunohistochemical analysis of S100A11 in surgically resected tissues from patients with MPM. The representative images of three MPM subtypes are shown. The pictures on the left show hematoxylin–eosin (HE)-stained images corresponding to the S100A11-stained images in the middle. Scale bars, 50 µm (positive and negative control) and 100 µm (clinical samples)
    Figure Legend Snippet: High secretion and overexpression of S100A11 in MPM. a Concentration of secreted S100A11 in the culture media of various cell lines, as determined by ELISA. The secretion level of S100A11 in all MPM cell lines, except for 211H, is higher than that of NM. 211H MSTO-211H, NM normal mesothelial cell, LC lung cancer, GC gastric cancer, CRC colorectal cancer, BC breast cancer. b Protein level of S100A11 was higher in MPM cell lines, compared to NM. NM normal mesothelial cell. c Immunohistochemical analysis of S100A11 in surgically resected tissues from patients with MPM. The representative images of three MPM subtypes are shown. The pictures on the left show hematoxylin–eosin (HE)-stained images corresponding to the S100A11-stained images in the middle. Scale bars, 50 µm (positive and negative control) and 100 µm (clinical samples)

    Techniques Used: Over Expression, Concentration Assay, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining, Negative Control

    32) Product Images from "Epigenetic downregulation of the DNA repair gene MED1/MBD4 in colorectal and ovarian cancer"

    Article Title: Epigenetic downregulation of the DNA repair gene MED1/MBD4 in colorectal and ovarian cancer

    Journal: Cancer biology & therapy

    doi:

    Relative expression of MED1 in adenomatous polyp or colorectal cancer specimens (N=39) compared to MED1 expression in matched normal mucosa from the same individual. (− = polyp, ● = stage I, ■ = stage II, ◆ = stage III, ▲= stage IV)
    Figure Legend Snippet: Relative expression of MED1 in adenomatous polyp or colorectal cancer specimens (N=39) compared to MED1 expression in matched normal mucosa from the same individual. (− = polyp, ● = stage I, ■ = stage II, ◆ = stage III, ▲= stage IV)

    Techniques Used: Expressing

    33) Product Images from "Insulin receptor isoform switching in intestinal stem cells, progenitors, differentiated lineages and tumors: evidence that IR-B limits proliferation"

    Article Title: Insulin receptor isoform switching in intestinal stem cells, progenitors, differentiated lineages and tumors: evidence that IR-B limits proliferation

    Journal: Journal of Cell Science

    doi: 10.1242/jcs.132985

    IR-B expression is enhanced in a non-transformed human intestinal epithelial cell line (HIECs) and Caco-2 cells with the capacity to spontaneously differentiate versus tumorigenic colorectal cancer (CRC) cell lines. RT-PCR using primers spanning exon
    Figure Legend Snippet: IR-B expression is enhanced in a non-transformed human intestinal epithelial cell line (HIECs) and Caco-2 cells with the capacity to spontaneously differentiate versus tumorigenic colorectal cancer (CRC) cell lines. RT-PCR using primers spanning exon

    Techniques Used: Expressing, Transformation Assay, Reverse Transcription Polymerase Chain Reaction

    34) Product Images from "Frequent Alteration of MLL3 Frameshift Mutations in Microsatellite Deficient Colorectal Cancer"

    Article Title: Frequent Alteration of MLL3 Frameshift Mutations in Microsatellite Deficient Colorectal Cancer

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0023320

    MLL3 mutation analysis in 8 colorectal cancer cell lines and 72 samples of primary colorectal tumors. (A) MLL3 has a poly-A(A) 9 tract within the coding sequence of exon 38. Homozygous frameshift mutations were found in RKO and HCT116, while heterozygous mutations were found in the microsatellite unstable cell lines SW48 and LoVo. Separate somatic mutations were found in SW48 and DLD1 c.10313G > A (p.G3438D), c.13630C > T (p.R4544W). (B) Heterozygous mutations were found in the same poly-A(A) 9 tract within the coding sequence of exon 38.
    Figure Legend Snippet: MLL3 mutation analysis in 8 colorectal cancer cell lines and 72 samples of primary colorectal tumors. (A) MLL3 has a poly-A(A) 9 tract within the coding sequence of exon 38. Homozygous frameshift mutations were found in RKO and HCT116, while heterozygous mutations were found in the microsatellite unstable cell lines SW48 and LoVo. Separate somatic mutations were found in SW48 and DLD1 c.10313G > A (p.G3438D), c.13630C > T (p.R4544W). (B) Heterozygous mutations were found in the same poly-A(A) 9 tract within the coding sequence of exon 38.

    Techniques Used: Mutagenesis, Sequencing

    35) Product Images from "Activation of the mTOR Pathway by Oxaliplatin in the Treatment of Colorectal Cancer Liver Metastasis"

    Article Title: Activation of the mTOR Pathway by Oxaliplatin in the Treatment of Colorectal Cancer Liver Metastasis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0169439

    Unsupervised hierarchical clustering on 39 colorectal cancer liver metastasis samples were performed with oxaliplatin treated samples boxed. Unsupervised hierarchical clustering showed that oxaliplatin treatment did not influence the clustering of the 39 samples.
    Figure Legend Snippet: Unsupervised hierarchical clustering on 39 colorectal cancer liver metastasis samples were performed with oxaliplatin treated samples boxed. Unsupervised hierarchical clustering showed that oxaliplatin treatment did not influence the clustering of the 39 samples.

    Techniques Used:

    Activation of the mTOR pathway by oxaliplatin in vivo . The level of phosphorylated p70 S6 kinase was increased in colorectal cancer PDXs by Day 2.
    Figure Legend Snippet: Activation of the mTOR pathway by oxaliplatin in vivo . The level of phosphorylated p70 S6 kinase was increased in colorectal cancer PDXs by Day 2.

    Techniques Used: Activation Assay, In Vivo

    Activation of the mTOR pathway by oxaliplatin in vitro . A. The level of phosphorylated p70 S6 kinase was increased in all five ATCC cell lines (HCT15, DLD-1, LoVo, HCT116, HT29 and Colo 205) by Day 3. B. The level of phosphorylated p70 S6 kinase was increased in all early passage colorectal cancer cell lines (CRC057, CRC119 and CRC240) by Day 3 or 4.
    Figure Legend Snippet: Activation of the mTOR pathway by oxaliplatin in vitro . A. The level of phosphorylated p70 S6 kinase was increased in all five ATCC cell lines (HCT15, DLD-1, LoVo, HCT116, HT29 and Colo 205) by Day 3. B. The level of phosphorylated p70 S6 kinase was increased in all early passage colorectal cancer cell lines (CRC057, CRC119 and CRC240) by Day 3 or 4.

    Techniques Used: Activation Assay, In Vitro

    Oxaliplatin and Rapamycin Synergy Graphs were performed to determine the combination index (CI). A . The growth of ATCC colorectal cell lines HCT15, DLD-1, LoVo, HCT116, HT29 and Colo 205 in the presence of oxaliplatin and rapamycin as single agents and in combination was analyzed by cytotoxicity assays (see materials and methods ) to determine the dual-effect of the two agents on the cell lines. Mean Combination Index (CI) values (from three experiments) at 25%, 50%, 75% and 90% is plotted for each cell line. (CI
    Figure Legend Snippet: Oxaliplatin and Rapamycin Synergy Graphs were performed to determine the combination index (CI). A . The growth of ATCC colorectal cell lines HCT15, DLD-1, LoVo, HCT116, HT29 and Colo 205 in the presence of oxaliplatin and rapamycin as single agents and in combination was analyzed by cytotoxicity assays (see materials and methods ) to determine the dual-effect of the two agents on the cell lines. Mean Combination Index (CI) values (from three experiments) at 25%, 50%, 75% and 90% is plotted for each cell line. (CI

    Techniques Used:

    36) Product Images from "Knockdown of CUL4B inhibits proliferation and promotes apoptosis of colorectal cancer cells through suppressing the Wnt/β-catenin signaling pathway"

    Article Title: Knockdown of CUL4B inhibits proliferation and promotes apoptosis of colorectal cancer cells through suppressing the Wnt/β-catenin signaling pathway

    Journal: International Journal of Clinical and Experimental Pathology

    doi:

    CUL4B is overexpressed in colorectal cancer cell lines. A. CUL4B mRNA expression was determined in several colorectal cancer cell lines and the human colon mucosa cell line. B. Western blot analysis was performed to examine CUL4B expression in colorectal
    Figure Legend Snippet: CUL4B is overexpressed in colorectal cancer cell lines. A. CUL4B mRNA expression was determined in several colorectal cancer cell lines and the human colon mucosa cell line. B. Western blot analysis was performed to examine CUL4B expression in colorectal

    Techniques Used: Expressing, Western Blot

    Knockdown of CUL4B promotes the apoptosis of colorectal cancer cells through Wnt/β-catenin signaling pathway. (A) The protein levels of β-catenin, cyclin D1 and c-Myc were determined by western blot analysis, β-actin was used as
    Figure Legend Snippet: Knockdown of CUL4B promotes the apoptosis of colorectal cancer cells through Wnt/β-catenin signaling pathway. (A) The protein levels of β-catenin, cyclin D1 and c-Myc were determined by western blot analysis, β-actin was used as

    Techniques Used: Western Blot

    37) Product Images from "Activation of the mTOR Pathway by Oxaliplatin in the Treatment of Colorectal Cancer Liver Metastasis"

    Article Title: Activation of the mTOR Pathway by Oxaliplatin in the Treatment of Colorectal Cancer Liver Metastasis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0169439

    Unsupervised hierarchical clustering on 39 colorectal cancer liver metastasis samples were performed with oxaliplatin treated samples boxed. Unsupervised hierarchical clustering showed that oxaliplatin treatment did not influence the clustering of the 39 samples.
    Figure Legend Snippet: Unsupervised hierarchical clustering on 39 colorectal cancer liver metastasis samples were performed with oxaliplatin treated samples boxed. Unsupervised hierarchical clustering showed that oxaliplatin treatment did not influence the clustering of the 39 samples.

    Techniques Used:

    Activation of the mTOR pathway by oxaliplatin in vivo . The level of phosphorylated p70 S6 kinase was increased in colorectal cancer PDXs by Day 2.
    Figure Legend Snippet: Activation of the mTOR pathway by oxaliplatin in vivo . The level of phosphorylated p70 S6 kinase was increased in colorectal cancer PDXs by Day 2.

    Techniques Used: Activation Assay, In Vivo

    Activation of the mTOR pathway by oxaliplatin in vitro . A. The level of phosphorylated p70 S6 kinase was increased in all five ATCC cell lines (HCT15, DLD-1, LoVo, HCT116, HT29 and Colo 205) by Day 3. B. The level of phosphorylated p70 S6 kinase was increased in all early passage colorectal cancer cell lines (CRC057, CRC119 and CRC240) by Day 3 or 4.
    Figure Legend Snippet: Activation of the mTOR pathway by oxaliplatin in vitro . A. The level of phosphorylated p70 S6 kinase was increased in all five ATCC cell lines (HCT15, DLD-1, LoVo, HCT116, HT29 and Colo 205) by Day 3. B. The level of phosphorylated p70 S6 kinase was increased in all early passage colorectal cancer cell lines (CRC057, CRC119 and CRC240) by Day 3 or 4.

    Techniques Used: Activation Assay, In Vitro

    Oxaliplatin and Rapamycin Synergy Graphs were performed to determine the combination index (CI). A . The growth of ATCC colorectal cell lines HCT15, DLD-1, LoVo, HCT116, HT29 and Colo 205 in the presence of oxaliplatin and rapamycin as single agents and in combination was analyzed by cytotoxicity assays (see materials and methods ) to determine the dual-effect of the two agents on the cell lines. Mean Combination Index (CI) values (from three experiments) at 25%, 50%, 75% and 90% is plotted for each cell line. (CI
    Figure Legend Snippet: Oxaliplatin and Rapamycin Synergy Graphs were performed to determine the combination index (CI). A . The growth of ATCC colorectal cell lines HCT15, DLD-1, LoVo, HCT116, HT29 and Colo 205 in the presence of oxaliplatin and rapamycin as single agents and in combination was analyzed by cytotoxicity assays (see materials and methods ) to determine the dual-effect of the two agents on the cell lines. Mean Combination Index (CI) values (from three experiments) at 25%, 50%, 75% and 90% is plotted for each cell line. (CI

    Techniques Used:

    38) Product Images from "Overexpression of NOX4 predicts poor prognosis and promotes tumor progression in human colorectal cancer"

    Article Title: Overexpression of NOX4 predicts poor prognosis and promotes tumor progression in human colorectal cancer

    Journal: Oncotarget

    doi: 10.18632/oncotarget.16829

    NOX4 expression is up-regulated in CRC ( A ) A meta-analysis of NOX4 gene expression from six Oncomine databases where colored squares indicate the median rank for NOX4 (vs. Normal tissue) across 10 analyses. Gaedcke Colorectal (1), Hong Colorectal (2), Kaiser Colon (3, 4), Skrzypczak Colorectal (5), Skrzypczak2 Colorectal (6), and TCGA (7–10). The P value is given for the median-rank analysis. ( B ) NOX4 mRNA expression was examined by RT-PCR and normalized to 18S expression ( P
    Figure Legend Snippet: NOX4 expression is up-regulated in CRC ( A ) A meta-analysis of NOX4 gene expression from six Oncomine databases where colored squares indicate the median rank for NOX4 (vs. Normal tissue) across 10 analyses. Gaedcke Colorectal (1), Hong Colorectal (2), Kaiser Colon (3, 4), Skrzypczak Colorectal (5), Skrzypczak2 Colorectal (6), and TCGA (7–10). The P value is given for the median-rank analysis. ( B ) NOX4 mRNA expression was examined by RT-PCR and normalized to 18S expression ( P

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    39) Product Images from "Modifying Effects of Glucose and Insulin/Insulin-Like Growth Factors on Colon Cancer Cells"

    Article Title: Modifying Effects of Glucose and Insulin/Insulin-Like Growth Factors on Colon Cancer Cells

    Journal: Frontiers in Oncology

    doi: 10.3389/fonc.2021.645732

    mRNA expression levels of the IGF-I (A) , IGF-II (B) , IGF-IR (C) , IGF-IIR (D) , > IR-A (E) and IR-B (F) (expressed as relative mRNA expression, normalized to the house-keeping genes HPRT , GUSB and ACTB ) in four human colorectal cancer cell lines. Two different conditions were tested: medium containing 5 mmol/L glucose (open bars) or 25 mmol/L glucose (black bars). Values represent the mean ± SEM of two independent experiments. *p
    Figure Legend Snippet: mRNA expression levels of the IGF-I (A) , IGF-II (B) , IGF-IR (C) , IGF-IIR (D) , > IR-A (E) and IR-B (F) (expressed as relative mRNA expression, normalized to the house-keeping genes HPRT , GUSB and ACTB ) in four human colorectal cancer cell lines. Two different conditions were tested: medium containing 5 mmol/L glucose (open bars) or 25 mmol/L glucose (black bars). Values represent the mean ± SEM of two independent experiments. *p

    Techniques Used: Expressing

    mRNA expression levels of the IGFBP-1 (A) , IGFBP-2 (B) , IGFBP-3 (C) and IGFBP-6 (D) (expressed as relative mRNA expression, normalized to the house-keeping genes HPRT, GUSB and ACTB ) in four human colorectal cancer cell lines. Two different conditions were tested: medium containing 5 mmol/L glucose (open bars) or 25 mmol/L glucose (black bars). Values represent the mean ± SEM of two independent experiments. *p
    Figure Legend Snippet: mRNA expression levels of the IGFBP-1 (A) , IGFBP-2 (B) , IGFBP-3 (C) and IGFBP-6 (D) (expressed as relative mRNA expression, normalized to the house-keeping genes HPRT, GUSB and ACTB ) in four human colorectal cancer cell lines. Two different conditions were tested: medium containing 5 mmol/L glucose (open bars) or 25 mmol/L glucose (black bars). Values represent the mean ± SEM of two independent experiments. *p

    Techniques Used: Expressing

    40) Product Images from "GPR126 regulates colorectal cancer cell proliferation by mediating HDAC2 and GLI2 expression, et al. GPR126 regulates colorectal cancer cell proliferation by mediating HDAC2 and GLI2 expression"

    Article Title: GPR126 regulates colorectal cancer cell proliferation by mediating HDAC2 and GLI2 expression, et al. GPR126 regulates colorectal cancer cell proliferation by mediating HDAC2 and GLI2 expression

    Journal: Cancer Science

    doi: 10.1111/cas.14868

    GPR126 knockdown attenuates colorectal cancer cell growth in vivo. A‐C, GPR126 knockdown in HT‐29, HCT116, and LoVo cells inhibited tumorigenesis in nude mice xenografts. Top panels, pictures of five representative tumors grown from each cell line. Bottom panels, statistical analysis of the volumes of 10 tumors formed from HCT‐116 (A), 15 tumors from HT‐29 (B), and eight tumors from LoVo (C). The volume curves were calculated on day 29 for HCT‐116 tumors (A), day 27 for HT‐29 (B), and day 35 for LoVo (C). Values are presented as the mean ± SEM. *** P
    Figure Legend Snippet: GPR126 knockdown attenuates colorectal cancer cell growth in vivo. A‐C, GPR126 knockdown in HT‐29, HCT116, and LoVo cells inhibited tumorigenesis in nude mice xenografts. Top panels, pictures of five representative tumors grown from each cell line. Bottom panels, statistical analysis of the volumes of 10 tumors formed from HCT‐116 (A), 15 tumors from HT‐29 (B), and eight tumors from LoVo (C). The volume curves were calculated on day 29 for HCT‐116 tumors (A), day 27 for HT‐29 (B), and day 35 for LoVo (C). Values are presented as the mean ± SEM. *** P

    Techniques Used: In Vivo, Mouse Assay

    GLI2 binding with HDAC2 to the PTCH1 promoter is regulated by HDAC2 and its expression recovery restores cell viability in GPR126‐silenced cells. A, Western blotting of HDAC2 and GLI2 protein levels in NC‐ and HDAC2‐shRNA‐infected HT‐29 cells. β‐actin was used as a loading control. B, Chromatin from HT‐29 cells was immunoprecipitated with anti‐HDAC2 antibodies and then eluted and immunoprecipitated again with normal mouse IgG(IgG) or anti‐Gli2(GLI2 Ab). Eluted DNA was PCR‐amplified using PTCH1 and GAPDH primers (B1). Quantitative PCR(qPCR) was used to examine the abundance of eluted DNA from B1, with the DNAs from input control as the internal control (B2). C, Ectopic expression of HDAC2 in GPR126‐silenced cells restores transcription activity of GLI2 promoter (Gli2‐Luc). NC or Sh1, cell infected by control virus or GPR126 shRNA virus; pGL3, empty luciferase vector; Gli2‐Luc, luciferase vector with Gli2 promoter; zs‐HDAC2, HDAC2 ectopic expression construct; zs, empty vector for cloning HDAC2 expression vector; ‘‐’, not used in co‐transfection; ‘+’, used in co‐transfection. D, E, the effect of Purmorphamine (hedgehog agonist) and GANT61 (GLI2 inhibitor), at indicated concentration, on cell viability and GLI2 protein expression in indicate colorectal cancer cells. DMSO, the solvent of Purmorphamine and GANT61. β‐actin was used as a loading control. F, Diagram illustrating the putative mechanisms of GPR126 function in colorectal cancer cells. GPR126 regulates SMO, GLI1, and HDAC2 expression ( GSE106696 ). GLI1 and GLI2, as the downstream components of GPR126 signaling, regulate the expression of hedgehog (HH) target genes, including PTCH1 , regulating tumor growth. HDAC2 is regulated by GPR126 regulated HDAC2, mediating GLI2 expression, and binds with GLI2 to the PTCH1 promoter or other promoters of HH target genes. The dotted line indicating ‘Not reported in this article’; solid line indicating ‘Has support in this article’; ‘?’ meaning ‘through unknown mechanism’
    Figure Legend Snippet: GLI2 binding with HDAC2 to the PTCH1 promoter is regulated by HDAC2 and its expression recovery restores cell viability in GPR126‐silenced cells. A, Western blotting of HDAC2 and GLI2 protein levels in NC‐ and HDAC2‐shRNA‐infected HT‐29 cells. β‐actin was used as a loading control. B, Chromatin from HT‐29 cells was immunoprecipitated with anti‐HDAC2 antibodies and then eluted and immunoprecipitated again with normal mouse IgG(IgG) or anti‐Gli2(GLI2 Ab). Eluted DNA was PCR‐amplified using PTCH1 and GAPDH primers (B1). Quantitative PCR(qPCR) was used to examine the abundance of eluted DNA from B1, with the DNAs from input control as the internal control (B2). C, Ectopic expression of HDAC2 in GPR126‐silenced cells restores transcription activity of GLI2 promoter (Gli2‐Luc). NC or Sh1, cell infected by control virus or GPR126 shRNA virus; pGL3, empty luciferase vector; Gli2‐Luc, luciferase vector with Gli2 promoter; zs‐HDAC2, HDAC2 ectopic expression construct; zs, empty vector for cloning HDAC2 expression vector; ‘‐’, not used in co‐transfection; ‘+’, used in co‐transfection. D, E, the effect of Purmorphamine (hedgehog agonist) and GANT61 (GLI2 inhibitor), at indicated concentration, on cell viability and GLI2 protein expression in indicate colorectal cancer cells. DMSO, the solvent of Purmorphamine and GANT61. β‐actin was used as a loading control. F, Diagram illustrating the putative mechanisms of GPR126 function in colorectal cancer cells. GPR126 regulates SMO, GLI1, and HDAC2 expression ( GSE106696 ). GLI1 and GLI2, as the downstream components of GPR126 signaling, regulate the expression of hedgehog (HH) target genes, including PTCH1 , regulating tumor growth. HDAC2 is regulated by GPR126 regulated HDAC2, mediating GLI2 expression, and binds with GLI2 to the PTCH1 promoter or other promoters of HH target genes. The dotted line indicating ‘Not reported in this article’; solid line indicating ‘Has support in this article’; ‘?’ meaning ‘through unknown mechanism’

    Techniques Used: Binding Assay, Expressing, Western Blot, shRNA, Infection, Immunoprecipitation, Polymerase Chain Reaction, Amplification, Activity Assay, Luciferase, Plasmid Preparation, Construct, Clone Assay, Cotransfection, Concentration Assay

    GPR126 is enriched in colorectal cancer cell lines and clinical colorectal cancer tissues. A, Relative GPR126 mRNA and protein expression levels evaluated by qRT‐PCR and western blotting, with the β‐actin as the internal control, in the indicated colorectal cancer cell lines. B, Representative immunohistochemistry staining of GPR126 (brown) in matched human colorectal cancer tissue and adjacent normal tissue. Scale bar = 50 µm. C, Statistical analysis of immunohistochemistry staining scores of GPR126 from 50 pairs of human colorectal cancer tissue (green) and matched adjacent normal colorectal tissue (grey) in the tissue microarray. The number on each column top represents the percentage of tissues per indicated grade. Chi square test was used for comparisons. *, P
    Figure Legend Snippet: GPR126 is enriched in colorectal cancer cell lines and clinical colorectal cancer tissues. A, Relative GPR126 mRNA and protein expression levels evaluated by qRT‐PCR and western blotting, with the β‐actin as the internal control, in the indicated colorectal cancer cell lines. B, Representative immunohistochemistry staining of GPR126 (brown) in matched human colorectal cancer tissue and adjacent normal tissue. Scale bar = 50 µm. C, Statistical analysis of immunohistochemistry staining scores of GPR126 from 50 pairs of human colorectal cancer tissue (green) and matched adjacent normal colorectal tissue (grey) in the tissue microarray. The number on each column top represents the percentage of tissues per indicated grade. Chi square test was used for comparisons. *, P

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Staining, Microarray

    GPR126 knockdown attenuates colorectal cancer cell growth in vitro. A‐C, Cell viability and growth of the indicated cells after GPR126 knockdown. The effect of two shRNAs targeting GPR126 (sh1, sh2) on GPR126 mRNA expression normalized to β‐actin mRNA in HT‐29 cells (A), cell viability of the indicated cells (B), and the growth of HT‐29 cells (C). NC, control vector. D, E, Cell colony formation in soft agar after shRNA (sh1)‐mediated GPR126 knockdown. NC, control vector. Representative images of the stained cell colonies (triplicate plates for each indicated cell line were used) (D). Quantification of cell colony numbers in (D) represented as a bar plot (E). Values are presented as the mean ± SEM. *** P
    Figure Legend Snippet: GPR126 knockdown attenuates colorectal cancer cell growth in vitro. A‐C, Cell viability and growth of the indicated cells after GPR126 knockdown. The effect of two shRNAs targeting GPR126 (sh1, sh2) on GPR126 mRNA expression normalized to β‐actin mRNA in HT‐29 cells (A), cell viability of the indicated cells (B), and the growth of HT‐29 cells (C). NC, control vector. D, E, Cell colony formation in soft agar after shRNA (sh1)‐mediated GPR126 knockdown. NC, control vector. Representative images of the stained cell colonies (triplicate plates for each indicated cell line were used) (D). Quantification of cell colony numbers in (D) represented as a bar plot (E). Values are presented as the mean ± SEM. *** P

    Techniques Used: In Vitro, Expressing, Plasmid Preparation, shRNA, Staining

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  • 86
    ATCC colorectal cancer cell lines
    Expression of PRDX2 is elevated in primary <t>colorectal</t> tumors compared with human colorectal tumor-adjacent tissues ( A ) Expression of PRDX2 protein in each of the primary colorectal tumors and colorectal adjacent noncancerous tissues paired from the same patient by Western blotting, GAPDH was used as loading control. ( B ) Expression of PRDX2 mRNA in each of the primary colorectal tumors (T) and colorectal adjacent noncancerous tissues (ANT) paired from the same patient by quantitative real-time reverse transcription-PCR (qRT-PCR). ( C ) PRDX2 expression level was up-regulated in the primary colorectal tumor compared with the paired colorectal adjacent noncancerous tissues from the same patient, as examined by immunohistochemistry. Data represent the mean ± SD of three experiments. (* p
    Colorectal Cancer Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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
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    86
    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.
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    Expression of PRDX2 is elevated in primary colorectal tumors compared with human colorectal tumor-adjacent tissues ( A ) Expression of PRDX2 protein in each of the primary colorectal tumors and colorectal adjacent noncancerous tissues paired from the same patient by Western blotting, GAPDH was used as loading control. ( B ) Expression of PRDX2 mRNA in each of the primary colorectal tumors (T) and colorectal adjacent noncancerous tissues (ANT) paired from the same patient by quantitative real-time reverse transcription-PCR (qRT-PCR). ( C ) PRDX2 expression level was up-regulated in the primary colorectal tumor compared with the paired colorectal adjacent noncancerous tissues from the same patient, as examined by immunohistochemistry. Data represent the mean ± SD of three experiments. (* p

    Journal: Oncotarget

    Article Title: Peroxiredoxin 2 is associated with colorectal cancer progression and poor survival of patients

    doi: 10.18632/oncotarget.14801

    Figure Lengend Snippet: Expression of PRDX2 is elevated in primary colorectal tumors compared with human colorectal tumor-adjacent tissues ( A ) Expression of PRDX2 protein in each of the primary colorectal tumors and colorectal adjacent noncancerous tissues paired from the same patient by Western blotting, GAPDH was used as loading control. ( B ) Expression of PRDX2 mRNA in each of the primary colorectal tumors (T) and colorectal adjacent noncancerous tissues (ANT) paired from the same patient by quantitative real-time reverse transcription-PCR (qRT-PCR). ( C ) PRDX2 expression level was up-regulated in the primary colorectal tumor compared with the paired colorectal adjacent noncancerous tissues from the same patient, as examined by immunohistochemistry. Data represent the mean ± SD of three experiments. (* p

    Article Snippet: The colorectal cancer cell lines, including HCT116, HT29, SW480, SW620, and SW837, were obtained from the American Type Culture Collection and kept in our laboratory.

    Techniques: Expressing, Western Blot, Polymerase Chain Reaction, Quantitative RT-PCR, Immunohistochemistry

    Expression analysis of PRDX2 protein and mRNA in HCEC and colorectal cancer cell lines by quantitative real-time reverse transcription-PCR (qRT-PCR) and Western blotting ( A ) Expression of PRDX2 protein in HCEC and cultured colorectal cancer cell lines SW480, SW620, HT29, HCT116, and SW837. ( B ) Expression of PRDX2 mRNA in HCEC and cultured colorectal cancer cell lines. Data represent the mean ± SD of three experiments. (* p

    Journal: Oncotarget

    Article Title: Peroxiredoxin 2 is associated with colorectal cancer progression and poor survival of patients

    doi: 10.18632/oncotarget.14801

    Figure Lengend Snippet: Expression analysis of PRDX2 protein and mRNA in HCEC and colorectal cancer cell lines by quantitative real-time reverse transcription-PCR (qRT-PCR) and Western blotting ( A ) Expression of PRDX2 protein in HCEC and cultured colorectal cancer cell lines SW480, SW620, HT29, HCT116, and SW837. ( B ) Expression of PRDX2 mRNA in HCEC and cultured colorectal cancer cell lines. Data represent the mean ± SD of three experiments. (* p

    Article Snippet: The colorectal cancer cell lines, including HCT116, HT29, SW480, SW620, and SW837, were obtained from the American Type Culture Collection and kept in our laboratory.

    Techniques: Expressing, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Cell Culture

    PRDX2 is frequently up-regulated in CRC ( A ) Representative immunohistochemical expression patterns of PRDX2 in 226 paired human primary colorectal cancer tissues and corresponding adjacent normal mucosa specimens are shown. ( B ) Percentage of cases with different staining intensity of PRDX2 in the tumor or adjacent normal tissues in the study cohort.

    Journal: Oncotarget

    Article Title: Peroxiredoxin 2 is associated with colorectal cancer progression and poor survival of patients

    doi: 10.18632/oncotarget.14801

    Figure Lengend Snippet: PRDX2 is frequently up-regulated in CRC ( A ) Representative immunohistochemical expression patterns of PRDX2 in 226 paired human primary colorectal cancer tissues and corresponding adjacent normal mucosa specimens are shown. ( B ) Percentage of cases with different staining intensity of PRDX2 in the tumor or adjacent normal tissues in the study cohort.

    Article Snippet: The colorectal cancer cell lines, including HCT116, HT29, SW480, SW620, and SW837, were obtained from the American Type Culture Collection and kept in our laboratory.

    Techniques: Immunohistochemistry, Expressing, Staining

    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