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

Figure 4 MiR-31-5p targets SATB2, and this targeting relationship is functionally relevant in ulcerative colitis and colorectal cancer. A: Venn diagram showing shared hsa-miR-31-5p target genes predicted by miRTarBase, miRWalk, and TargetScan; B: Quantitative reverse transcription polymerase chain reaction analysis of SATB2 and PPP2R2A mRNA levels in HCT116 cells transfected with miR-31-5p mimic or inhibitor. ^aP < 0.05 vs control, ^bP < 0.01 vs control (n = 3); C: Schematic representation of wild-type and mutant luciferase reporter constructs containing the miR-31-5p binding site in the 3’-untranslated region of SATB2; D: Dual-luciferase reporter assay confirming that miR-31-5p directly targeted SATB2 but not PPP2R2A. ^bP < 0.01 vs NC + SATB2-wild-type (n = 3); E: Western blot analysis of SATB2 protein expression in HCT116 cells following miR-31-5p modulation. ^aP < 0.05 vs control, ^bP < 0.01 vs control (n = 4); F-H: Heatmap, volcano plot, and box plot showing differentially expressed mRNAs and SATB2 downregulation in human ulcerative colitis inflamed vs adjacent control tissues. ^aP < 0.05 vs N (n = 4 pairs); I: Correlation analysis between miR-31-5p and SATB2 expression in human ulcerative colitis tissues, r = -0.9; J-L: Validation of SATB2 expression in HCT116 colorectal cancer cells and dextran sulfate sodium-induced colitis in miR-31 transgenic mice by quantitative reverse transcription polymerase chain reaction, western blot, and immunohistochemistry. J, ^bP < 0.01 vs NCM-460; K and L: ^aP < 0.05 vs control, ^bP < 0.01 vs control, ^dP < 0.05 vs dextran sulfate sodium, ^eP < 0.01 vs dextran sulfate sodium; M: Kyoto Encyclopedia of Genes and Genomes and Gene Ontology enrichment analyses of SATB2-associated downstream genes. KEGG: Kyoto Encyclopedia of Genes and Genomes; GO: Gene Ontology; DSS: Dextran sulfate sodium; UC: Ulcerative colitis.