Affect of miR-195 on proliferation and apoptosis of glioma U251 cells

Abstract

Objective To explore the function and mechanism of miR-195 over-expression in glioblastoma U251 cell proliferation and apoptosis.Methods The experiment is divided into Blank,NC and miR-195.Blank is treated with equal volume PBS.Negative control or miR-195 mimics were transfected into U251 ceils by Lipofectamine RNAiMAX.miR-195 expression levels were detected by Real -time PCR.Cell growth inhibition rates and proliferation were analyzed by CCK-8 assay and colony formation.Flow cytometry was used to monitor changes in cell apoptosis and cycle.Apoptotic cells and necrotic cells were also qualified by Hoechst 33342 and propidium iodide staining.The target of miR-195 was forecasted by bioinformatics.The expression level of Bcl-2 protein was detected through Western blotting.Results After the over-expression of miR-195,Real-time PCR showed that miR-195 expression level was up-regulated about six times.Cell growth inhibition rate and colony formation revealed that the growth and proliferation ability of the miR-195 group were significantly suppressed(compared with NC,P ＜0.001).Over-expression of miR-195 could promote cell apoptosis and death(compared with Blank and NC,P ＜ 0.001).Predicting outcomes of TargetScan indicated that Bcl-2 was a target of miR 195.Furthermore,the over-expression of miR-195 could block G0/G1 transition and dowm-regulate the Bcl-2 expression (compared with Blank and NC,P ＜ 0.05).Conclusion Over-expression of miR-195 could inhibit proliferation,induce apoptosis and death,and block G0/S1 transition.miR-195 may serve as an attractive target of gene therapy for glioblastoma. Key words: Glioma; MiR-195 ; Proliferation; Apoptosis; Bcl-2