Resveratrol suppresses ulcerative colitis and colitis-associated colorectal cancer via modulating the miR-31-5p/SATB2 axis

Figure 7 Resveratrol suppresses malignant phenotypes of HCT116 cells via the miR-31-5p/SATB2 axis. A: CCK-8 assay of HCT116 cell viability after 24 hours of exposure to Resveratrol (Res) (0, 2.5, 5, 10, 20, and 40 μg/mL). ^aP < 0.05 vs 0 μg/mL, ^bP < 0.01 vs 0 μg/mL (n = 8); B: Quantitative reverse transcription polymerase chain reaction analysis of miR-31-5p in HCT116 cells treated with 10 μg/mL Res for 0, 12, or 24 hours. ^bP < 0.01 vs 0 hours (n = 3); C: Quantitative reverse transcription polymerase chain reaction quantification of miR-31 expression in colonic tissues from colitis-associated colorectal cancer mice with Res intervention. ^aP < 0.05 vs control, ^bP < 0.01 vs control, ^eP < 0.01 vs azoxymethane/dextran sulfate sodium (n = 3); D: Western blots of SATB2 protein expression in HCT116 cells treated with Res (0, 2.5, 5, 10, 20, and 40 μg/mL). ^bP < 0.01 vs control (n = 3); E and F: Cell migration evaluated by scratch wound assays under miR-31-5p overexpression or inhibition with Res treatment (scale bar = 200 μm). ^aP < 0.05 vs control, ^bP < 0.01 vs control, ^eP < 0.01 vs miR-31-5p mimic, ^gP < 0.05 vs miR-31-5p inhibitor (n = 3); G: Colony formation assay evaluating the impact of Res on miR-31-5p-modulated proliferative capacity. ^aP < 0.05 vs control, ^bP < 0.01 vs control, ^dP < 0.05 vs miR-31-5p mimic, ^gP < 0.05 vs miR-31-5p inhibitor (n = 3); H: Flow cytometric analysis of apoptosis in HCT116 with altered miR-31-5p expression with/without Res treatment. ^bP < 0.01 vs control, ^eP < 0.01 vs miR-31-5p mimic, ^hP < 0.01 vs miR-31-5p inhibitor (n = 3); I: Western blots of SATB2 and nuclear factor-κB expression and the Bcl-2/Bax ratio after miR-31-5p manipulation with/without Res treatment. ^aP < 0.05 vs control, ^bP < 0.01 vs control, ^dP < 0.05 vs miR-31-5p mimic, ^eP < 0.01 vs miR-31-5p mimic, ^hP < 0.01 vs miR-31-5p inhibitor (n = 3); J and K: Scratch wound assays with SATB2-silenced (si-SATB2) or SATB2-overexpressing (OE-SATB2) HCT116 cells treated with Res (scale bar = 200 μm). ^aP < 0.05 vs control, ^bP < 0.01 vs control, ^eP < 0.01 vs si-SATB2, ^gP < 0.05 vs OE-SATB2 (n = 3); L and M: Colony formation assays with si-SATB2 or OE-SATB2 cells with Res treatment. ^aP < 0.05 vs control, ^bP < 0.01 vs control, ^eP < 0.01 vs si-SATB2, ^gP < 0.05 vs OE-SATB2 (n = 3); N and O: Western blots of apoptosis-related protein expression levels in si-SATB2 or OE-SATB2 cells following Res treatment. ^aP < 0.05 vs control, ^bP < 0.01 vs control, ^dP < 0.05 vs si-SATB2, ^eP < 0.01 vs si-SATB2, ^gP < 0.05 vs OE-SATB2, ^hP < 0.01 vs OE-SATB2 (n = 3). Res: Resveratrol; NS: Not significant; NF-κB: Nuclear factor-κB; AOM: Azoxymethane; DSS: Dextran sulfate sodium; Cyt-C: Cytochrome C.