Down-regulation of miR-30a-3p/5p promotes esophageal squamous cell carcinoma cell proliferation by activating the Wnt signaling pathway

Figure 6 miR-30a-3p/5p directly target the 3'-UTRs of Wnt2 and Fzd2, respectively. A: Target gene screening of miR-30a-3p (left) and miR-30a-5p (right) by qPCR suggested that miR-30a-3p targeted Wnt2 and miR-30a-5p targeted Fzd2 in the Wnt signaling pathway; B: Predicted target sequences of miR-30a-3p (top) and miR-30a-5p (bottom) in the 3’-UTRs of WNT2 and FZD2, respectively; C: qPCR (top) and Western blot (bottom), respectively, confirmed the inhibitory effects of miR-30a-3p and miR-30a-5p on Wnt2 and Fzd2 at both mRNA and protein levels. GAPDH and α-tubulin were, respectively, used as loading controls for qPCR and Western blot; D: Luciferase reporter assay indicated that miR-30-3p and miR-30a-5p directly targeted the wild-type 3’-UTRs of Wnt2 and Fzd2, respectively, but had no effect on the mutant ones. Two concentrations of miR-30a-3p- and miR-30a-5p-mimics (10 nmol/L and 20 nmol/L) plus wild-type or mutant 3’-UTRs of target genes were applied. Error bars represent the mean ± SD of three independent experiments; ^aP < 0.05.