Journal: Cancer Research

Article Title: Colorectal Cancer Cell Lines Are Representative Models of the Main Molecular Subtypes of Primary Cancer

doi: 10.1158/0008-5472.can-14-0013

Figure Lengend Snippet: Figure 5. Overlap of mutations and DNA copy-number states between paired colorectal cancer cell lines. COLO201/COLO205, CX-1/HT29, Gp2D/Gp5D, LS174T/LS180, and DLD1/HCT8/HCT15 were derived from the same tumor, SW480/SW620 from a primary tumor and subsequent lymph node metastasis.

Article Snippet: Bodmer laboratory, CACO2, COLO201, COLO205, COLO320-DM, DLD1, HCC2998, HCT116, HCT15, HCT8, HT29, LOVO, LS174T, LS180, LS411, LS513, NCI-H716, NCI-H747, RKO, SKCO-1, SNU-C1, SNU-C2B, SW1116, SW1417, SW1463, SW403, SW48, SW480, SW620, SW837, SW948, and T84 were obtained from the American Type Culture Collection, KM12 was obtained from DCTD, COLO678, and CX-1 were obtained from DSMZ, Gp2D, Gp5D, HT115, and HT55 were obtained from ECACC, CCK81, and CoCM-1 were obtained from HSRRB, VACO10, and VACO4S were provided by Dr. J.A.

Techniques: Derivative Assay

Journal: Cell Proliferation

Article Title: SATB 2 targeted by methylated miR‐34c‐5p suppresses proliferation and metastasis attenuating the epithelial‐mesenchymal transition in colorectal cancer

doi: 10.1111/cpr.12455

Figure Lengend Snippet: SATB2 prevents metastasis of colorectal cancer (CRC) cells in vivo. A, Bioluminescent images of in vivo incidence of lung metastasis and primary tumour in nude mice subjected to tail vein inoculation with HT‐29 cells whose endogenous SATB2 was stably knocked down (labelled as HT‐29 Lv‐sh‐SATB2) or not (HT‐29 Lv‐control). Fourteen nude mice were grouped into 2, with each having 7 mice. The incidence rate of lung metastasis in group where mice inoculated with HT‐29 Lv‐sh‐SATB2 cells was 6 of 7. In contrast, the incidence of lung metastasis in group where mice inoculated with HT‐29 Lv‐control cells were 1 of 7; there was significant difference with P value being .029 using Cross‐table analysis. B, Haematoxylin‐eosin (HE) section analysis of lung metastasis nodes and its quantitative counts of visible nodes per lung; ***P < .001 compared with Lv‐control group using the independent sample t‐test analysis. C, Similarly and parallel to HT‐29 cells, the evaluation of incidence of lung metastasis was verified in another kind of CRC cell line HCT‐15 whose endogenous SATB2 was stably knocked down (HCT‐15 Lv‐sh‐SATB2) or not (HCT‐15 Lv‐control). As presented, there was also significant difference in the case of incidence of lung metastasis, with P value being .021 using cross‐table analysis. D, HE section analysis of lung metastasis nodes and its quantitative counts of visible nodes per lung, ***P < .001 relative to Lv‐control group using the independent sample t‐test analysis. In terms of bioluminescent image presented, shown were the representative figures selected among the 7 candidates

Article Snippet: The human CRC cell lines HT‐29, Colo‐320, SW480, SW620 and HCT‐15 were obtained from China Center for Type Culture Collection.

Techniques: In Vivo, Stable Transfection

Journal: Cell Proliferation

Article Title: SATB 2 targeted by methylated miR‐34c‐5p suppresses proliferation and metastasis attenuating the epithelial‐mesenchymal transition in colorectal cancer

doi: 10.1111/cpr.12455

Figure Lengend Snippet: Repressed miR‐34c‐5p in CRC resulted from methylation in its promoter sequence. A, Basal level of miR‐34c‐5p in the same panel of CRC cell lines as SATB2 was detected using immunoblotting. B, Detection of miR‐34c‐5p level in the 85 paired cases of CRC and its normal control using qRT‐PCR. U6, the internal loading control. Independent sample t test was used to analyse the statistical difference between CRC and its paired normal control. C, Spearman correlation was carried out to analyse the correlation between miR‐34c‐5p and SATB2 expression on mRNA level. As displayed, the R 2 coefficient was .131, the sample size was 85 cases and P value was .0007. D, Detection of methylated miR‐34c‐5p level in CRC and paired normal control using methylated qRT‐PCR. E, Similarly, Spearman correlation was carried out to analyse the correlation between methylated miR‐34c‐5p and miR‐34c‐5p expression on mRNA level, as shown, the R 2 coefficient was −.401, the sample size was 85 cases, and P value was .003. F, the miR‐34c‐5p level was assayed before and after treatment with 5‐aza‐2′‐deoxycytidine (abbreviated as 5‐Aza‐dC), an inhibitor for DNA methyltransferase, for 48 h in CRC cell lines HT‐29 and HCT‐15. *P < .05, **P < .01, ***P < .001 in comparison with their control group, that is, HCoEpiC using independent sample t test. G: expression variation of SATB2 as well as E‐cadherin and N‐cadherin in the presence and absence of 5‐Aza‐dC treatment for 48 h, as exemplified by western blot

Article Snippet: The human CRC cell lines HT‐29, Colo‐320, SW480, SW620 and HCT‐15 were obtained from China Center for Type Culture Collection.

Techniques: Methylation, Sequencing, Western Blot, Quantitative RT-PCR, Expressing

Journal: Cell Proliferation

Article Title: SATB 2 targeted by methylated miR‐34c‐5p suppresses proliferation and metastasis attenuating the epithelial‐mesenchymal transition in colorectal cancer

doi: 10.1111/cpr.12455

Figure Lengend Snippet: MiR‐34c‐5p was identified to be able to directly and negatively modulate SATB2. A, Bioinformatic prediction of the potential binding sites of SATB2 in the mature sequence of miR‐34c‐5p, which was highly conserved in mammalians. The bold blue words were the highly conserved binding site of miR‐34c‐5p binding to SATB2. B, Verification of the potential binding of miR‐34c‐5p with SATB2 using Luciferase reporter assay. Independent sample t test was used to analyse the statistical difference relative to control groups. Experiment was performed in triplicate and shown was the representative. C, Expression variation of SATB2 as well as the typical EMT relevant biomarkers, including E‐cadherin and N‐cadherin, after transfection with miR‐34c‐5p mimics and inhibitor sequence into HT‐29 and SW480 cells, as exemplified by immunoblot. The experiment was performed independently in triplicate and shown were the representative figures selected among the candidates. D, In vivo evaluation of metastatic variation of CRC cells whose endogenous miR‐34c‐5p was stably re‐expressed using lentivirus (Lv) vector fused with bioluminescent tag. For HT‐29 cells, 14 nude mice were grouped into 2, with each 7. In control group, mice were subjected to the tail vein inoculation of HT‐29 cells transfected with Lv‐miR‐34c‐5p‐scramble (hereafter referred to as Lv‐control), whereas in experimental group, mice subjected to the tail vein inoculation of HT‐29 cells transfected with Lv‐miR‐34c‐5p. The same holds true for HCT‐15 cells. Presented were the representative bioluminescent images picked up among the candidates from at least 3 different times of repeat. *P<0.05 compared with control group using independent sample t test

Article Snippet: The human CRC cell lines HT‐29, Colo‐320, SW480, SW620 and HCT‐15 were obtained from China Center for Type Culture Collection.

Techniques: Binding Assay, Sequencing, Luciferase, Reporter Assay, Expressing, Transfection, Western Blot, In Vivo, Stable Transfection, Plasmid Preparation

Journal: Cell Proliferation

Article Title: SATB 2 targeted by methylated miR‐34c‐5p suppresses proliferation and metastasis attenuating the epithelial‐mesenchymal transition in colorectal cancer

doi: 10.1111/cpr.12455

Figure Lengend Snippet: SATB2 suppresses the epithelial‐mesenchymal transition (EMT) in colorectal cancer (CRC) cell lines. A, Endogenous basal level of SATB2 in a panel of CRC cell lines available to us; the observed band size of SATB2 and β‐actin band was 83 and 42 kilodalton (kDa), respectively; B, transient knock‐down of SATB2 using siRNA technique in HT‐29 and HCT‐15 cells. Scramble‐siRNA was used as negative control of siRNA sequences transfected. The observed band size of E‐cadherin, N‐cadherin and vimentin band was 110, 125 and 54 kDa, respectively; C, Likewise, exogenous SATB2 fused with green fluorescent protein (GFP) was re‐expressed in CRC cell lines HT‐29 and HCT‐15. The observed band size of SATB2 fused with GFP was approximately 110 kDa, with GFP tag being observed at 27 kDa. Shown were the representative figures picked among the candidates from at least independently 3 different times of repeat

Article Snippet: The human CRC cell lines HT‐29, Colo‐320, SW480, SW620 and HCT‐15 were obtained from China Center for Type Culture Collection.

Techniques: Negative Control, Transfection

Journal: EJNMMI Research

Article Title: Indium-111-labeled CD166-targeted peptide as a potential nuclear imaging agent for detecting colorectal cancer stem-like cells in a xenograft mouse model

doi: 10.1186/s13550-020-0597-3

Figure Lengend Snippet: Isolation and analysis of CD166 + HCT15 cells. a Schematic diagram for the isolation of CD166 + HCT15 cells by magnetic-activated cell sorting (MACS). b The rate of CD166 expression in HCT15 cell sorted population determined by flow cytometry. Both CD166 + HCT15 and CD166 − HCT15 cells were treated with IgG-FITC and FITC-labeled CD166 antibody (CD166ab-FITC, 20 μg/mL) for 1 h. Data are presented as mean ± SD ( n ≥ 3). * P < 0.05, versus IgG-FITC. c The protein levels of CD166 in both CD166 + HCT15 and CD166 − HCT15 cells. The protein expression was detected by Western blotting and quantified by densitometry and normalized by GAPDH levels. The data are presented as mean ± SD ( n ≥ 3). * P < 0.05, versus CD166 − HCT15

Article Snippet: The human CRC-derived cell line (HCT15) was obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Isolation, FACS, Expressing, Flow Cytometry, Labeling, Western Blot

Journal: EJNMMI Research

Article Title: Indium-111-labeled CD166-targeted peptide as a potential nuclear imaging agent for detecting colorectal cancer stem-like cells in a xenograft mouse model

doi: 10.1186/s13550-020-0597-3

Figure Lengend Snippet: Identification of cancer stem cell feature in CD166 + HCT15 cells. a Morphological characteristics during the growth of tumor spheres for 14 days. The quantitation of cell sphere formation was detected by manual calculation under light microscopy. Magnification, × 400; scale bar, 50 μm. b Colony formation of CD166 + HCT15 and CD166 − HCT15 cells. The representative image was shown and the colony number was quantified. c Cell migration assay. A wound was made in the monolayer in both CD166 + HCT15 and CD166 − HCT15 cells (T0), and then the cells were allowed to migrate for 24 h (T24). The quantification of wound closure was determined by ImageJ software. Magnification, ×100; scale bar, 200 μm. In a – c , data are presented as mean ± SD ( n = 3).* P < 0.05, versus CD166 − group

Article Snippet: The human CRC-derived cell line (HCT15) was obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Quantitation Assay, Light Microscopy, Cell Migration Assay, Software

Journal: EJNMMI Research

Article Title: Indium-111-labeled CD166-targeted peptide as a potential nuclear imaging agent for detecting colorectal cancer stem-like cells in a xenograft mouse model

doi: 10.1186/s13550-020-0597-3

Figure Lengend Snippet: The protein expressions of cancer stem cell markers and drug resistance in CD166 + HCT15 cells. a Four cancer stem cell markers Nanog, c-Myc, OCT4, and Survivin were detected by Western blotting and quantified by densitometry and normalized by GAPDH levels. The data are presented as mean ± SD ( n ≥ 3). * P < 0.05, versus CD166 − HCT15. The arrow indicated the protein position. b The effects of doxorubicin and 5-fluouracil on cell viability. Both CD166 + HCT15 and CD166 − HCT15 cells were treated with doxorubicin and 5-fluouracil (1–50 μM) for 24 h. Data are presented as mean ± SD ( n ≥ 5). * P < 0.05, versus CD166 − HCT15. # P < 0.05, versus Ctrl (control)

Article Snippet: The human CRC-derived cell line (HCT15) was obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Western Blot, Control

Journal: EJNMMI Research

Article Title: Indium-111-labeled CD166-targeted peptide as a potential nuclear imaging agent for detecting colorectal cancer stem-like cells in a xenograft mouse model

doi: 10.1186/s13550-020-0597-3

Figure Lengend Snippet: Screening and identification of CD166-targeted peptide. The phage display (12 phage library) was used to screen CD166 binding phages with four rounds of biopanning. a The titers of recovered phages from each round of biopanning were eluted by a blue/white colony screening on LB/IPTG/Xgal plate. The phage enrichment rate was calculated as output number/input number. b Phage ELISA assay. The carrying different peptide sequences of phage clones were exposed to CD166 protein and BSA (as a control) for detecting the binding affinity. c Phage binding to CD166 expression cells. The phage clones were further screened by cell-based phage ELISA. The CD166 + HCT15 and CD166 − HCT15 cells were positive and negative CD166 protein expression cells, respectively. Data are presented as mean ± SD ( n ≥ 3)

Article Snippet: The human CRC-derived cell line (HCT15) was obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Binding Assay, Colony Assay, Enzyme-linked Immunosorbent Assay, Clone Assay, Control, Expressing

Journal: EJNMMI Research

Article Title: Indium-111-labeled CD166-targeted peptide as a potential nuclear imaging agent for detecting colorectal cancer stem-like cells in a xenograft mouse model

doi: 10.1186/s13550-020-0597-3

Figure Lengend Snippet: CD166tp-G 18 C targeted to CD166-positive CRC cells. a The CD166tp-G 18 C binding assay in CRC cells was analyzed by flow cytometry. Both CD166 + HCT15 and CD166 − HCT15 cells were treated with CD166tp-G 18 C-FITC (20 μg/mL) for 1 h. In competitive group, CD166 + HCT15 cells were pre-treated with CD166tp-G 18 C (20 μg/mL) for 1 h and then treated with 20 μg/mL CD166tp-G 18 C-FITC for 1 h. Data are presented as mean ± SD ( n ≥ 3). * P < 0.05, versus (CD166 − HCT15) CD166tp-G 18 C-FITC; # P < 0.05, versus (CD166 + HCT15) CD166tp-G 18 C-FITC. b The CD166tp-G18C binding assay in both CD166 + HCT15 and CD166 − HCT15 cells was analyzed by fluorescent microscopy. Both cells were treated with CD166tp-G 18 C-FITC (20 μg/mL) for 2 h. The total nuclei were stained with 4’, 6-diamino-2-phenylindole (DAPI). Magnification, × 200; scale bar, 100 μm. c CD166tp-G 18 C and CD166 protein interaction assay. Both CD166 + HCT15 and CD166 − HCT15 cells were treated with CD166tp-G 18 C-FITC (20 μg/mL) for 1 h. The supernatant of lysates was collected and treated with or without biotinylated-CD166 polyclonal antibody (2 μg/mL) in the presence of streptavidin agarose beads at 4 °C overnight. The fluorescent signaling of immunoprecipitates was analyzed by ELISA. The CD166tp-G 18 C (20 μg/mL) was pre-treated for competitive inhibition assay. Data are presented as mean ± SD ( n ≥ 3). * P < 0.05

Article Snippet: The human CRC-derived cell line (HCT15) was obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Binding Assay, Flow Cytometry, Microscopy, Staining, Protein Interaction Assay, Enzyme-linked Immunosorbent Assay, Inhibition

Journal: EJNMMI Research

Article Title: Indium-111-labeled CD166-targeted peptide as a potential nuclear imaging agent for detecting colorectal cancer stem-like cells in a xenograft mouse model

doi: 10.1186/s13550-020-0597-3

Figure Lengend Snippet: The CD166 imaging in tumors and organs of CRC xenograft mice. a The CD166 imaging of both CD166 + HCT15 and CD166 − HCT15 xenograft model. Both CD166 + HCT15 and CD166 − HCT15 cells were subcutaneously inoculated into the right flank of nude mice for establishing CD166 high- and non-expressed xenograft model, respectively. The Cy5.5-conjugated CD166tp-G 18 C was intravenously injected into both xenograft mice for 6 h and observed by in vivo imaging system. b The distribution of CD166 imaging in various organs and tumor tissue of CD166 + HCT15 xenograft mice. The top panel presented the CD166 imaging in various organs. The button panel presented the quantitation of fluorescent signals. Data are presented as mean ± SD ( n = 3). * P < 0.05, versus other organs. c The protein expressions of CD166 in tumors and various organs of CD166 + HCT15 xenograft mice. d The CD166 expressions in tumors of both CD166 + HCT15 and CD166 − HCT15 xenograft mice. In c and d , protein expressions were determined by Western blotting and quantified by densitometry and normalized by GAPDH levels. In c , data are presented as mean ± SD ( n = 3). * P < 0.05, versus other organs. In d , data are presented as mean ± SD ( n = 3). * P < 0.05, versus 100 mm 3 (CD166 − HCT15); # P < 0.05, versus 150 mm 3 (CD166 − HCT15)

Article Snippet: The human CRC-derived cell line (HCT15) was obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Imaging, Injection, In Vivo Imaging, Quantitation Assay, Western Blot

Journal: EJNMMI Research

Article Title: Indium-111-labeled CD166-targeted peptide as a potential nuclear imaging agent for detecting colorectal cancer stem-like cells in a xenograft mouse model

doi: 10.1186/s13550-020-0597-3

Figure Lengend Snippet: The synthesis and stability assay for 111 In-DTPA-CD166tp-G 18 C. a The flowchart of 111 In-DTPA-CD166tp-G 18 C synthesis. CD166tp-G 18 C was conjugated with DTPA for indium-111 labeling and then intravenously injected into CD166 + HCT15 xenograft mice for CD166 detection in tumor area. b The molecular weights of DTPA-CD166tp-G 18 C detection. The molecular weights of Maleimide-DTPA, CD166-G 18 C, and DTPA-CD166tp-G 18 C were confirmed by a mass spectrometry. c The labeling efficiency of 111 In-DTPA-CD166tp-G 18 C. After indium-111 labeling, instant thin-layer chromatography (ITLC) was performed to determine the radio-labeling efficiency in 111 In-DTPA-CD166tp-G 18 C. d The stability of 111 In-DTPA-CD166tp-G 18 C. The radioactivity of 111 In-DTPA-CD166tp-G 18 C was detected in human, fetal bovine, and mouse serum for 144 h at 37 °C. The radioactivity was detected by ITLC. Data are presented as mean ± SD ( n = 5)

Article Snippet: The human CRC-derived cell line (HCT15) was obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Stability Assay, Labeling, Injection, Mass Spectrometry, Thin Layer Chromatography, Radioactivity

Journal: EJNMMI Research

Article Title: Indium-111-labeled CD166-targeted peptide as a potential nuclear imaging agent for detecting colorectal cancer stem-like cells in a xenograft mouse model

doi: 10.1186/s13550-020-0597-3

Figure Lengend Snippet: The nuclear imaging tracer of 111 In-DTPA-CD166tp-G 18 C for detection of CD166-positive colorectal tumor in vivo. a The colorectal tumor nuclear imaging analysis in CD166 + HCT15 xenograft mice. The 111 In-DTPA-CD166tp-G 18 C and control groups (740 MBq/kg/per mouse) were intravenously injected into mice for 2, 4, 24, and 48 h and detected by a nanoSPECT/CT. Group I, 111 In-DTPA; Group II, 111 In-DTPA-G 18 C; Group III, 111 In-DTPA-CD166tp-C; Group IV, 111 In-DTPA-CD166tp-G 18 C. b Quantification of nuclear images in tumor areas of colorectal tumor xenograft mice. The circled positions in images were quantified by a 3D analysis software. Data are presented as mean ± SD ( n ≥ 3). * P < 0.05, versus control group. c The competitive study of 111 In-DTPA-CD166tp-G 18 C in CD166 + HCT15 xenograft mice. After tumor xenograft mice were intravenously injected with CD166tp-G 18 C (0, 10, and 50 mg/kg) for 6 h, 111 In-DTPA-CD166tp-G 18 C (740 MBq/kg/mouse) was intravenously injected into mice for 24 and 48 h and detected by a nanoSPECT/CT. d Quantification of nuclear images in tumor areas of colorectal tumor xenograft mice. Data are presented as mean ± SD ( n ≥ 3). * P < 0.05, versus 0 mg/kg CD166tp-G 18 C group, # P < 0.05, versus 0 mg/kg CD166tp-G 18 C group

Article Snippet: The human CRC-derived cell line (HCT15) was obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Imaging, In Vivo, Control, Injection, Software

Journal: EJNMMI Research

Article Title: Indium-111-labeled CD166-targeted peptide as a potential nuclear imaging agent for detecting colorectal cancer stem-like cells in a xenograft mouse model

doi: 10.1186/s13550-020-0597-3

Figure Lengend Snippet: Tissue bio-distribution of 111 In-DTPA-CD166tp-G 18 C in colorectal tumor xenograft mice. After 111 In-DTPA-CD166tp-G 18 C and control groups ( 111 In-DTPA, 111 In-DTPA-G 18 C and 111 In-DTPA-CD166tp-C) (148 MBq/kg/per mouse) intravenous injection for 2, 4, 24, and 48 h, CD166 + HCT15 xenograft mice were sacrificed and obtained tumor, blood, and organs. The radioactivity of samples was detected by a gamma counter. Values are presented as the percentage of injected dose per gram organ (ID%/g). Data are presented as mean ± SD ( n ≥ 3). * P < 0.05, versus control group

Article Snippet: The human CRC-derived cell line (HCT15) was obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Control, Injection, Radioactivity

Journal: Frontiers in Cell and Developmental Biology

Article Title: CircFAM120B Blocks the Development of Colorectal Cancer by Activating TGF-Beta Receptor II Expression via Targeting miR-645

doi: 10.3389/fcell.2021.682543

Figure Lengend Snippet: CircFAM120B was downregulated in colorectal cancer (CRC) tissues and cells. (A,B) A microarray profile revealed that circFAM120B was a differently expressed circRNA that was significantly downregulated in CRC tissues ( n = 10). (C) The expression of circFAM120B in tumor tissues ( n = 50) and normal tissues ( n = 50) was detected by quantitative real-time polymerase chain reaction (qRT-PCR). (D) The expression of circFAM120B in LoVo, HCT15, and NCM460 cells was detected by qRT-PCR. (E,F) LoVo and HCT15 cells were exposed to actinomycin D to test the stability of circFAM120B. * P < 0.05.

Article Snippet: Colorectal cancer cell lines, including LoVo and HCT15, were purchased from ProCell Co., Ltd. (Wuhan, China) and maintained in matched special medium (ProCell Co., Ltd.).

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

Journal: Frontiers in Cell and Developmental Biology

Article Title: CircFAM120B Blocks the Development of Colorectal Cancer by Activating TGF-Beta Receptor II Expression via Targeting miR-645

doi: 10.3389/fcell.2021.682543

Figure Lengend Snippet: CircFAM120B overexpression weakened cell proliferation, migration/invasion and glycolysis. In circFAM120B-overexpressed LoVo and HCT15 cells, (A–C) cell proliferation was assessed by CCK-8 assay and colony formation assay. (D,E) Cell migration and invasion were determined by transwell assay (magnification: 100×). (F,G) Extracellular acidification rate (ECAR) was measured by glycolysis stress test to evaluate glycolysis metabolism. (H,I) Lactate production and glucose consumption were quantified to assess glycolysis progression using corresponding kits. (J,K) Glycolysis-related markers, HK2 and LDHA, were quantified by western blot to observe glycolysis. * P < 0.05.

Article Snippet: Colorectal cancer cell lines, including LoVo and HCT15, were purchased from ProCell Co., Ltd. (Wuhan, China) and maintained in matched special medium (ProCell Co., Ltd.).

Techniques: Over Expression, Migration, CCK-8 Assay, Colony Assay, Transwell Assay, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: CircFAM120B Blocks the Development of Colorectal Cancer by Activating TGF-Beta Receptor II Expression via Targeting miR-645

doi: 10.3389/fcell.2021.682543

Figure Lengend Snippet: MiR-645 was a target of circFAM120B. (A,B) Biotinylated-circFAM120B probe could significantly enrich the level of miR-645. (C) Wild-type circFAM120B sequence containing miR-645 binding sites was mutated to generate mutant-type circFAM120B sequence for dual-luciferase reporter assay. (D,E) Dual-luciferase reporter assay was performed to further validate the relationship between miR-645 and circFAM120B. (F) RIP assay was performed to validate the relationship between miR-645 and circFAM120B. (G) The expression of miR-645 in tumor tissues ( n = 50) and normal tissues ( n = 50) was detected by qRT-PCR. (H) The expression of miR-645 in LoVo, HCT15, and NCM460 cells was detected by qRT-PCR. (I) The expression of miR-645 in LoVo and HCT15 cells with circFAM120B overexpression was detected by qRT-PCR. * P < 0.05.

Article Snippet: Colorectal cancer cell lines, including LoVo and HCT15, were purchased from ProCell Co., Ltd. (Wuhan, China) and maintained in matched special medium (ProCell Co., Ltd.).

Techniques: Sequencing, Binding Assay, Mutagenesis, Luciferase, Reporter Assay, Expressing, Quantitative RT-PCR, Over Expression

Journal: Frontiers in Cell and Developmental Biology

Article Title: CircFAM120B Blocks the Development of Colorectal Cancer by Activating TGF-Beta Receptor II Expression via Targeting miR-645

doi: 10.3389/fcell.2021.682543

Figure Lengend Snippet: CircFAM120B blocked CRC progression in vitro by targeting miR-645. LoVo and HCT15 cells were transfected with oe-circFAM120B alone or oe-circFAM120B + miR-645, with vector or oe-circFAM120B + miR-NC as a control. (A) The expression of miR-645 in these transfected cells was detected by qRT-PCR. (B,C) Cell proliferation was assessed by CCK-8 assay. (D) Cell proliferation was assessed by colony formation assay. (E,F) Cell migration and invasion were monitored by transwell assay (magnification: 100×). (G,H) Glycolysis was evaluated by ECAR using glycolysis stress test. (I,J) Lactate production and glucose consumption were quantified to monitor glycolysis. (K,L) The expression of HK2 and LDHA was quantified by western blot. * P < 0.05.

Article Snippet: Colorectal cancer cell lines, including LoVo and HCT15, were purchased from ProCell Co., Ltd. (Wuhan, China) and maintained in matched special medium (ProCell Co., Ltd.).

Techniques: In Vitro, Transfection, Plasmid Preparation, Expressing, Quantitative RT-PCR, CCK-8 Assay, Colony Assay, Migration, Transwell Assay, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: CircFAM120B Blocks the Development of Colorectal Cancer by Activating TGF-Beta Receptor II Expression via Targeting miR-645

doi: 10.3389/fcell.2021.682543

Figure Lengend Snippet: TGFBR2 was a target of miR-645. (A) Wild-type TGFBR2 3′UTR sequence containing miR-645 binding sites was mutated to generate mutant-type TGFBR2 3′UTR sequence for dual-luciferase reporter assay. (B,C) Dual-luciferase reporter assay was conducted to verify the interaction between miR-645 and TGFBR2. (D) RIP assay was conducted to verify the interaction between miR-645 and TGFBR2. (E,F) The expression of TGFBR2 suppressed by miR-645 restoration was detected by qRT-PCR and western blot. (G,H) The expression of TGFBR2 in tumor tissues ( n = 50) and normal tissues ( n = 50) was detected by qRT-PCR and western blot. (I,J) The expression of TGFBR2 in LoVo, HCT15 and NCM460 cells was detected by qRT-PCR and western blot. * P < 0.05.

Article Snippet: Colorectal cancer cell lines, including LoVo and HCT15, were purchased from ProCell Co., Ltd. (Wuhan, China) and maintained in matched special medium (ProCell Co., Ltd.).

Techniques: Sequencing, Binding Assay, Mutagenesis, Luciferase, Reporter Assay, Expressing, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: CircFAM120B Blocks the Development of Colorectal Cancer by Activating TGF-Beta Receptor II Expression via Targeting miR-645

doi: 10.3389/fcell.2021.682543

Figure Lengend Snippet: miR-645 promoted CRC progression in vitro by mediating TGFBR2. LoVo and HCT15 cells were transfected with anti-miR-645 or anti-miR-645 + si-TGFBR2, with anti-miR-NC or anti-miR-645 + si-NC as the corresponding control. (A,B) The expression of TGFBR2 was examined using qRT-PCR and western blot. (C–E) Cell proliferation was investigated using CCK-8 assay and colony formation assay. (F,G) Cell migration and invasion were monitored by transwell assay (magnification: 100×). (H,I) ECAR was measured by glycolysis stress test. (J,K) Lactate production and glucose consumption were detected using the corresponding kits. (L,M) The expression of HK2 and LDHA was measured by western blot. * P < 0.05.

Article Snippet: Colorectal cancer cell lines, including LoVo and HCT15, were purchased from ProCell Co., Ltd. (Wuhan, China) and maintained in matched special medium (ProCell Co., Ltd.).

Techniques: In Vitro, Transfection, Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Colony Assay, Migration, Transwell Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: CircFAM120B Blocks the Development of Colorectal Cancer by Activating TGF-Beta Receptor II Expression via Targeting miR-645

doi: 10.3389/fcell.2021.682543

Figure Lengend Snippet: CircFAM120B activated the expression of TGFBR2 by targeting miR-645. In LoVo and HCT15 cells transfected with oe-circFAM120B, vector, oe-circFAM120B + miR-645 or oe-circFAM120B + miR-NC, the expression of TGFBR2 was detected by (A,B) qRT-PCR and (C) western blot. * P < 0.05.

Article Snippet: Colorectal cancer cell lines, including LoVo and HCT15, were purchased from ProCell Co., Ltd. (Wuhan, China) and maintained in matched special medium (ProCell Co., Ltd.).

Techniques: Expressing, Transfection, Plasmid Preparation, Quantitative RT-PCR, Western Blot

Journal: PLoS ONE

Article Title: A Novel Microtubule Inhibitor 4SC-207 with Anti-Proliferative Activity in Taxane-Resistant Cells

doi: 10.1371/journal.pone.0079594

Figure Lengend Snippet: HCT-15 Xenograft model.

Article Snippet: The HCT-15 xenograft model was performed at VivoPharm (Kenttown, Australia).

Techniques: Control