Effects of microRNA-483-3p on human glioma cells / 中国病理生理杂志

ABSTRACT

AIM: To investigate the effects of microRNA (miRNA)-483-3p on the growth and migration of human glioma cell line A172 and its potential mechanisms.METHODS: The abundance of miRNA-483-3p in human embryonic kidney 293 cells and different human glioma cell lines (A172, U251 and SHG44) was measured by RT-qPCR.After down-regulation of miRNA-483-3p by transfection of inhibitor in the A172 cells, the cell viability, cell cycle distribution and cell migration were detected by CCK-8 assay, flow cytometry and Transwell assay, respectively.Furthermore, the protein levels of cell cycle-related molecules and epithelial-mesenchymal transition markers were measured by Western blot.Luciferase reporter assay was used to predict and verify the target gene of miRNA-483-3p.RESULTS: miRNA-483-3p was highly expressed in human glioma cells.Knockdown of miRNA-483-3p inhibited A172 cell viability, arrested cell cycle and decreased cell migration rate.Furthermore, the protein levels of cyclin D1, cyclin-dependent kinase 4, phoshorylated retinoblastoma protein, N-cadherin and vimentin were significantly decreased after knockdown of miRNA-483-3p, accompanied with the up-regulation of E-cadherin and β-catenin protein expression.Luciferase reporter assay demonstrated that Smad4 was a potential target gene of miRNA-483-3p.Down-regulation of Smad4 in the A172 cells transfected with miRNA-483-3p inhibitor partially reversed the effect of miRNA-483-3p on cell viability and migration.CONCLUSION: Knockdown of miRNA-483-3p restrains the growth and migration of A172 cells by targeting Smad4.