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: 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