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

Figure 5 SATB2 partially counteracts miR-31-5p-driven malignant phenotypes in HCT116 cells. A: Quantitative reverse transcription polymerase chain reaction identification of the optimal si-SATB2 concentration in HCT116 cells. ^bP < 0.01 vs control, ^eP < 0.01 vs si-SATB2 (3 nM), ^hP < 0.01 vs si-SATB2 (5 nM) (n = 3); B: Western blot validation of SATB2 overexpression efficiency following 200 nM OE-SATB2 plasmid transfection. ^bP < 0.01 vs control (n = 4); C: Wound healing assays showing partial reversal of miR-31-5p-enhanced HCT116 cell migration by SATB2, (scale bar = 200 μm). ^aP < 0.05 vs NC + vector, ^dP < 0.05 vs vector + mimic, ^gP < 0.05 vs NC + OE-SATB2 (n = 3); D: Colony formation assays demonstrating attenuation of miR-31-5p-induced HCT116 cell proliferation by SATB2. ^bP < 0.01 vs NC + vector, ^dP < 0.05 vs vector + mimic, ^hP < 0.01 vs NC + OE-SATB2 (n = 3); E: Flow cytometric analysis of HCT116 cells apoptosis indicating partial rescue of miR-31-5p-mediated anti-apoptotic effects by SATB2. ^bP < 0.01 vs NC + vector, ^eP < 0.01 vs vector + mimic, ^hP < 0.01 vs NC + OE-SATB2 (n = 3); F: Western blotting of inflammatory signaling proteins (Toll-like receptor 4, nuclear factor-κB, cyclooxygenase-2, tumor necrosis factor-α) following miR-31-5p and SATB2 modulation in HCT116 cells. ^aP < 0.05 vs NC + vector, ^bP < 0.01 vs NC + vector, ^dP < 0.05 vs vector + mimic, ^eP < 0.01 vs vector + mimic, ^gP < 0.05 vs NC + OE-SATB2, ^hP < 0.01 vs NC + OE-SATB2 (n = 3); G: Western blots of mitochondrial apoptosis-related proteins (Bcl-2, Bax, cytochrome C, caspase-9, caspase-3) assessing SATB2-mediated reversal of miR-31-5p anti-apoptotic signaling in HCT116 cells. ^aP < 0.05 vs NC + vector, ^bP < 0.01 vs NC + vector, ^dP < 0.05 vs vector + mimic, ^eP < 0.01 vs vector + mimic, ^gP < 0.05 vs NC + OE-SATB2, ^hP < 0.01 vs NC + OE-SATB2 (n = 3). TLR-4: Toll-like receptor 4; NF-κB: Nuclear factor-κB; COX-2: Cyclooxygenase-2; TNF-α: Tumor necrosis factor-α; Cyt-C: Cytochrome C.