Journal: Oncotarget

Article Title: BRG1 targeting STAT3/VEGFC signaling regulates lymphangiogenesis in colorectal cancer

doi: 10.18632/oncotarget.9038

Figure Lengend Snippet: A. Expression of BRG1 protein and VEGFC mRNA in 8 cultured CRC cell lines (HT29, HCT116, LoVo, Caco-2, KM12, SW48, SW480 and SW620). B. (Left) Analyses showing the expression of BRG1 protein and VEGFC mRNA in 10 freshly human CRC tissues. (Right) BRG1 protein expression was negatively correlated with VEGFC mRNA expression in 10 freshly human CRC tissues. C. (Left) BRG1 levels were negatively correlated with VEGFC expression in primary human CRC specimens ( n =31). Micrographs of BRG1 and VEGFC expression level in human CRC tissues. Original magnification, ×100. (Right) Percentage of CRC specimens with low or high BRG1 expression relative to VEGFC expression. * p <0.05.

Article Snippet: All CRC cell lines (LoVo, SW480, HT29, HCT116, Caco-2, KM12, SW48 and SW620) and the HEK293T cell line were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Cell Culture

Journal: Molecular Therapy Oncology

Article Title: Aurora kinase A drives non-canonical YAP1/TAZ crosstalk to sustain primary resistance to anti-EGFR therapies in colorectal cancer

doi: 10.1016/j.omton.2025.201032

Figure Lengend Snippet: TAZ overexpression sustains YAP1-dependent cetuximab resistance (A) Western blot illustrating the levels of TAZ, YAP1, and YAP1-activating phosphorylation at Ser397 ( n = 3). (B) Western blot illustrating the levels of MET ( n = 3), TAZ ( n = 3), YAP1 ( n = 3), and p-YAP1(Ser397) ( n = 4) after YAP1 knockdown in the SW48 cell line. Results are plotted as the mean ± SD. Differences between groups were studied using a two-tailed t test. (C) Western blot illustrating protein expression of MET ( n = 3), p-MET ( n = 4), TAZ ( n = 3), YAP1 ( n = 3), and p-YAP(Ser397) ( n = 4) after silencing YAP1 or TAZ in the C10 cell line. Results are plotted as the mean ± SD. Differences between groups were studied using an one-way ANOVA test followed by Tukey’s test for multiple comparisons. (D) Cell proliferation analysis of SW48 cell line after silencing YAP1 ( n = 3). Results are plotted as the mean ± SD. Differences between groups were studied using a two-tailed t test. ∗∗ p < 0.01. (E) Cell proliferation analysis of C10 cell line after silencing YAP1 or TAZ ( n = 3). Results are plotted as the mean ± SD. Differences between groups were studied using a two-tailed t test. ∗ p < 0.05.

Article Snippet: SW48 and NCIH-508 were obtained from the American Type Culture Collection.

Techniques: Over Expression, Western Blot, Phospho-proteomics, Knockdown, Two Tailed Test, Expressing

Journal: Molecular Therapy Oncology

Article Title: Aurora kinase A drives non-canonical YAP1/TAZ crosstalk to sustain primary resistance to anti-EGFR therapies in colorectal cancer

doi: 10.1016/j.omton.2025.201032

Figure Lengend Snippet: TAZ sustains YAP1-driven acquisition of stemness traits in CRC cell lines (A) Colony-formation assay after silencing of YAP1 or TAZ in the cetuximab-resistant SW48 and C10 cell lines. (B) ALDH1 activity assay performed in SW48 and C10 cell lines with knockdown of YAP1 or TAZ compared with the empty vector controls. (C) Spheroid formation assay of C10 after YAP1 or TAZ silencing. Spheroid volume and number were measured. (D) Western blot illustrating the levels of the stemness markers CD133 ( n = 4) and CD44 ( n = 3) in the C10 cell line after knocking-down YAP1, TAZ, or both, compared with cell line transduced with an empty vector. (E) Gene expression levels of the stemness marker POU5F1 in the C10 cell line in the C10 cell line after knocking-down YAP1, TAZ, or both compared with cell line transduced with an empty vector ( n = 4). Results are plotted as the mean ± SD. Differences between groups were studied using a two-tailed t test in SW48 cell line. A one-way ANOVA and Tukey's multiple comparisons test were performed to analyze data obtained from the C10 cell line. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: SW48 and NCIH-508 were obtained from the American Type Culture Collection.

Techniques: Colony Assay, Activity Assay, Knockdown, Plasmid Preparation, Tube Formation Assay, Western Blot, Transduction, Gene Expression, Marker, Two Tailed Test

Journal: Frontiers in Oncology

Article Title: Selective vulnerability of ARID1A deficient colon cancer cells to combined radiation and ATR-inhibitor therapy

doi: 10.3389/fonc.2022.999626

Figure Lengend Snippet: Effect of ATRi on radiosensitivity. (A) ARID1A expression in colon cancer lines; (B) Effect of ATRi (VE822) on radiosensitivity. ARID1A+ and ARID1A- cell lines were pre-treated for 1 h with 20 nM VE822 and irradiated with 0 Gy, 2 Gy, 4 Gy and 6 Gy. Plating efficiency of sham treated (untr) and VE822 treated (ATRi) cells were plotted as log10 for ARID1A + and ARID1A - cell lines. Results of 3 independent experiments are shown for CRC cell lines.

Article Snippet: The human CRC cell lines with mutant (LS180, RKO, SW48) and wild-type (HCT15, HCT116 and Colo320DM) ARID1A were obtained from ATCC (LGC Standards, Wesel, Germany) and were designated as ARID1A - and ARID1A + cells, respectively.

Techniques: Expressing, Irradiation

Journal: Frontiers in Oncology

Article Title: Selective vulnerability of ARID1A deficient colon cancer cells to combined radiation and ATR-inhibitor therapy

doi: 10.3389/fonc.2022.999626

Figure Lengend Snippet: Cell cycle effect of ATRi/ARID1A. (A) Synchronized cells in early S phase and mid S phase were pre-treated for 1 h with VE822 and irradiated thereafter with 0 Gy, 2 Gy and 4 Gy. Plating efficiency of sham treated (untr) and VE822 treated (ATRi) cells were plotted as log10 for ARID1A + and ARID1A - cell lines. (B) Exit from G2 phase into the M phase was measured after treatment with VE822 and irradiation in ARID1A - SW48 and ARID1A + HCT116 cell lines. Fraction of phospho-histone H3 positive cells were plotted against time after irradiation. MI = phospho-histone H3 in radiation/phospho-histone H3 in no-radiation × 100%. Results of 3 independent experiments are shown for CRC cell lines.

Article Snippet: The human CRC cell lines with mutant (LS180, RKO, SW48) and wild-type (HCT15, HCT116 and Colo320DM) ARID1A were obtained from ATCC (LGC Standards, Wesel, Germany) and were designated as ARID1A - and ARID1A + cells, respectively.

Techniques: Irradiation

Journal: Frontiers in Oncology

Article Title: Selective vulnerability of ARID1A deficient colon cancer cells to combined radiation and ATR-inhibitor therapy

doi: 10.3389/fonc.2022.999626

Figure Lengend Snippet: Effect of ATRi on ɣH2AX foci formation in G2-phase CRC cell lines.Maximum intensity projection (MIP) images of γH2AX foci (red) at tmax (1h) in G2-phase ARID1A + (A, B) and ARID1A - (C, D) cells without (untr) and with 20 nM VE822 (ATRi) in EdU - (green) cells after exposure to the indicated IR doses. Cells were counterstained with DAPI (blue). The respective numbers of γH2AX foci at tmax as a function of IR dose are shown in (B) (ARID1A + cells) and (D) (ARID1A - cells). Results of 3 independent experiments are shown for CRC cell lines.

Article Snippet: The human CRC cell lines with mutant (LS180, RKO, SW48) and wild-type (HCT15, HCT116 and Colo320DM) ARID1A were obtained from ATCC (LGC Standards, Wesel, Germany) and were designated as ARID1A - and ARID1A + cells, respectively.

Techniques:

Journal: Frontiers in Oncology

Article Title: Selective vulnerability of ARID1A deficient colon cancer cells to combined radiation and ATR-inhibitor therapy

doi: 10.3389/fonc.2022.999626

Figure Lengend Snippet: Effect of ATRi on RAD51 foci formation in G2-phase CRC cells. Maximum intensity projection (MIP) images of RAD51 foci (red) at tmax (6h) in G2-phase ARID1A+ (A, B) and ARID1A- (C, D) cells without (untr) and with 20 nM VE822 (ATRi) in EdU- (green) cells after exposure to the indicated IR doses. Cells were counterstained with DAPI (blue). The respective numbers of Rad51 foci at tmax as a function of IR dose are shown in (B) (ARID1A+ cells) and (D) (ARID1A- cells). Results of 3 independent experiments are shown for CRC cell lines.

Article Snippet: The human CRC cell lines with mutant (LS180, RKO, SW48) and wild-type (HCT15, HCT116 and Colo320DM) ARID1A were obtained from ATCC (LGC Standards, Wesel, Germany) and were designated as ARID1A - and ARID1A + cells, respectively.

Techniques:

Journal: Frontiers in Oncology

Article Title: Selective vulnerability of ARID1A deficient colon cancer cells to combined radiation and ATR-inhibitor therapy

doi: 10.3389/fonc.2022.999626

Figure Lengend Snippet: Effect of ATRi on HR repair in reporter cell lines. (A) Western blot results of ARID1A knock-down in DR-GFP-U2OS and DR-GFP-A549 reporter cells; GAPDH was used as an internal control. (B) Normalized GFP expression in DR-GFP- U2OS and DR-GFP-A549 reporter cells after treatment with control siRNA (mock), 20 nM VE822 (ATRi), ARID1A specific siRNA (siARID1A) and ATRi after knock-down of ARID1A (siARID1A, ATRi). (C) Normalized GFP expression in SA-GFP- U2OS reporter cells after treatment with control siRNA (mock), 20 nM VE822 (ATRi), ARID1A specific siRNA (siARID1A) and ATRi after knock-down of ARID1A (siARID1A, ATRi). (D) Normalized GFP expression in EJ2-GFP- U2OS reporter cells after treatment with control siRNA (mock), 20 nM VE822 (ATRi), ARID1A specific siRNA (siARID1A) and ATRi after knock-down of ARID1A (siARID1A, ATRi).Results of 3 independent experiments are shown for CRC cell lines.

Article Snippet: The human CRC cell lines with mutant (LS180, RKO, SW48) and wild-type (HCT15, HCT116 and Colo320DM) ARID1A were obtained from ATCC (LGC Standards, Wesel, Germany) and were designated as ARID1A - and ARID1A + cells, respectively.

Techniques: Western Blot, Knockdown, Control, Expressing

Journal: Frontiers in Oncology

Article Title: Selective vulnerability of ARID1A deficient colon cancer cells to combined radiation and ATR-inhibitor therapy

doi: 10.3389/fonc.2022.999626

Figure Lengend Snippet: Effect of ATRi on ex vivo explants from CRC patients. (A) Example of IHC staining of ARID1A expression from clinical CRC tumor cells with and without ARID1A expression. (B) Western blot of CK20 expression from primary CRC cells; GAPDH was used as an internal control. (C) ATP-Tumor Chemosensitivity Assay for the effect of the ATR inhibitor VE822 on ex-vivo cells from CRC patients. ATP activity was measured after treatment of ex-vivo cells with concentration ranged from 0, 2.5 up to 100 nM in cells from CRC patients with (+) and without (-) ARID1A expression.

Article Snippet: The human CRC cell lines with mutant (LS180, RKO, SW48) and wild-type (HCT15, HCT116 and Colo320DM) ARID1A were obtained from ATCC (LGC Standards, Wesel, Germany) and were designated as ARID1A - and ARID1A + cells, respectively.

Techniques: Ex Vivo, Immunohistochemistry, Expressing, Western Blot, Control, Activity Assay, Concentration Assay

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

Article Title: Metformin selectively inhibits metastatic colorectal cancer with the KRAS mutation by intracellular accumulation through silencing MATE1

doi: 10.1073/pnas.1918845117

Figure Lengend Snippet: Down-regulation of MATE1 under KRAS mutation is associated with the sensitivity of CRC cells to metformin. ( A – C ) Transcriptional levels of SLC29A4 (PMAT), SLC22A1 (OCT1), SLC22A2 (OCT2), SLC22A3 (OCT3), SLC47A1 (MATE1), and SLC47A2 (MATE2k) in KRAS WT CRC cell lines SW48 and CaCO2, KRAS G13D CRC cell lines HCT-116 and LoVo ( A ), KRAS G13D SW48 and its counterpart SW48 ( B ), and sh-KRAS LoVo and sh-ctrl LoVo ( C ) were determined by qRT-PCR ( n = 3). Data are shown as mean ± SEM. P values were determined with the black bar as a control in A – C as * P < 0.05, ** P < 0.01. Each analysis was replicated three times. ( D ) Representative images of MATE1 immunohistochemistry on cross-sections from patients with mCRC with T2DM and metformin use, and the different integral optical density (IOD) of MATE1 between the KRAS wild type and mutation group is shown. The correlation between MATE1 expression and Ki67 level of 27 mCRC patients with T2DM and metformin use was determined by Pearson’s correlation analysis. ( E and F ) Forty-eight-hour growth of the KRAS G13D CRC cell line LoVo with or without MATE1 overexpression ( E ) and KRAS WT CRC cell line SW48 with or without MATE1 knockdown by small interfering RNA ( F ) was detected after treatment with 2.5, 5, 10, and 20 mM metformin ( n = 5). Data are shown as mean ± SEM. The difference compared with the control group at each concentration was determined as ** P < 0.01. ( G – L ) The representative morphology ( G ), tumor growth rate, and tumor weight ( L ) are shown as a result of 30-d treatment with metformin in SW48 xenograft ( H ), SW48 with sh-MATE1 xenograft ( I ), KRAS G13D SW48 ( J ), and KRAS G13D SW48 with LV-MATE1 xenograft ( K ) models. Data are shown as mean ± SEM. All P values were determined by two-way ANOVA. ** P < 0.01.

Article Snippet: The human CRC cell lines (SW48, CaCO2, LoVo, HCT-116, HT-29) were obtained from the American Type Culture Collection.

Techniques: Mutagenesis, Quantitative RT-PCR, Control, Immunohistochemistry, Expressing, Over Expression, Knockdown, Small Interfering RNA, Concentration Assay

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

Article Title: Metformin selectively inhibits metastatic colorectal cancer with the KRAS mutation by intracellular accumulation through silencing MATE1

doi: 10.1073/pnas.1918845117

Figure Lengend Snippet: KRAS mutation down-regulates MATE1 through mediating the hypermethylation status on the CpG island of the MATE1 promoter. ( A ) The correlation between MATE1 transcriptional levels and methylation levels of CpG sites on the promoter of MATE1. Data from TCGA-COAD RNA-seq-HTseq-FPKM-521 (workflow type HTSeq-FPKM, normalized from RNA-seq of 521 samples) and methyArray 450k were conducted by Pearson’s correlation analysis. ( B ) Bisulfite sequencing PCR analysis showing the methylation status of CpG sites on the MATE1 promoter in CRC cell lines. ( C ) Immunoblot analysis of MATE1 levels and BSP analysis of the MATE1 promoter are shown between KRAS G13D SW48 and its counterpart SW48. ( D ) Immunoblot analysis monitoring MATE1 levels and CpG-site methylation on the MATE1 promoter in KRAS shRNA-LoVo cells treated with dimethyl sulfoxide (DMSO) as a negative control or azacitidine as a positive control. ( E ) The MATE1 expression in 374469 KRAS WT colon adenocarcinoma and 386650 KRAS G12D colon mucinous adenocarcinoma was analyzed by immunoblot, and the methylation status of CpG sites on the MATE1 promoter was determined by BSP. Data are shown as mean ± SEM. All P values were determined by two-way ANOVA. * P < 0.05, ** P < 0.01.

Article Snippet: The human CRC cell lines (SW48, CaCO2, LoVo, HCT-116, HT-29) were obtained from the American Type Culture Collection.

Techniques: Mutagenesis, Methylation, RNA Sequencing, Methylation Sequencing, Western Blot, shRNA, Negative Control, Positive Control, Expressing