MiR-32 activates PI3K-Akt pathway by targeting toPTENand enhances Runx2 expression and phosphorylation.
A, The miR-32 mimics (pre-miR32),negative control of miR-32 mimics (pre-con), miR-32 inhibitors (anti-miR32), or negative control of miR-32 inhibitors (anti-con)were transfected into MVSMCs and the cells were lysed and analyzed by immunobloting with the indicated antibody. *P< 0.05, **P< 0.01, ***P< 0.001, compared to control groups. B, Effect of miR-32 on PTEN expression by luciferase reporter assay. MVSMCs were cotransfected with pre-miR32 (or anti-miR32) and PTEN 3’UTR wild-type vectors (PTEN 3’UTR-WT) or PTEN 3’UTR mutant vectors (PTEN 3’UTR-MUT). Luciferase activity was normalized by the ratio of firefly and renilla luciferase signals. C, PI3K signaling is involved in miR-32 induced Runx2 expression and phosphorylation. MVSMCs were transfected with indicated concentration of miR-32 mimics or pre-con, with or without LY294002 and total protein extracts were analyzed by Western blot.Data of panel Aare analyzed with ANOVA with Bonferroni correction, and student t test is used for data anaysis of panel B&C. *P< 0.05,**P< 0.01.