miR-497-5p targets FZD4 and FZD6 and inhibits Wnt/β-catenin signaling pathway by decreasing the expression of FZD4 and FZD6.
Notes: (A) The putative miR-497-5p-binding sites in the 3′-UTR of FZD4 and FZD6 were predicted by TargetScan. (B) The binding of miR-497-5p and FZD4 or FZD6 was validated by dual-luciferase reporter assay. Unlike negative control (NC) mimics, miR-497-5p mimics reduced the luciferase activity of wild-type (WT) FDZ4 and FDZ6 reporter vectors, but not that of mutant (MUT) reporter vectors. (C) Overexpression of miR-497-5p decreased the expression of FZD4, FZD6, and β-catenin, while knockdown of miR-497-5p increased the expression of FZD4, FZD6, and β-catenin. (D) The overexpression efficiency of pcDNA3.1-FZD4 and pcDNA3.1-FZD6 was determined by qRT-PCR and Western blot. (E) miR-497-5p mimics decreased the expression of FZD4, FZD6, and β-catenin, while overexpression vectors of FZD4 or FZD6 abrogated the regulation effect of miR-497-5p on FZD4 or FZD6. The expression of β-catenin was abrogated by either overexpression vectors of FZD4 or FZD6. Data are presented as mean ± SD. ***P<0.001.
Abbreviation: qRT-PCR, quantitative reverse transcription PCR.