The Effect And Mechanism Of Down-regulating MiR-586 On Glioma Cell Proliferation And Cycle

Objective:Glioma is now considered a genetic and epigenetic disease with a poor prognosis.MicroRNA(miRNA)genetic variation is considered to be an important potential risk factor for gliomas.Methods:1.To detect the expression level of microRNA-586(miR-586)in clinical tissue samples of glioblastoma and non-tumor brain tissues were used real-time fluorescence quantitative PCR experiment,and to detect miR-586 in glioma cell lines and expression levels in human normal astrocytes were used same experiment;2.CCK-8,plate cloning,flow cytometry and EdU experiments were used to detect the biological effects of miRNA-586 inhibition on U87 cells;3.To predicting the downstream target gene of miR-586 through the miRNA target gene online prediction website,and verifying the binding site of miR-586 at the 3’-UTR of the downstream target gene through the dual luciferase reporter gene experiment to confirm whether SFRP1 was direct target of miR-586,Western blotting test was used to detect the effect of miR-586 on the expression of downstream target gene SFRP1;4.Co-transfected si-SFRP1 in the U87 cell line that down-regulated miR-586,and tested whether the inhibition of SFRP1 can attenuate the biological effects of down-regulated miR-586 on U87 cells,and used CCK-8,flow cytometry,EdU experiments and Western blotting experiment for analysis;5.After U87 cells with miR-586 down-regulated were implanted into the lateral ventricle of nude mice,Kaplan-meier survival analysis,hematoxylin-eosin staining,and immunohistochemistry were used to analyze the effect of miR-586 suppression on glioma in vivo.Results:1.MiR-586 was highly expressed in glioma tissues and cells,and significantly expressed in U87 cells;2.Inhibiting the expression of miR-586 can significantly inhibit the proliferation of glioma cells and induced glioma cell arrest in G0/G1 phase;3.SFRP1 was the downstream target gene of miR-586,and inhibited the expression of miR-586 can upregulate the expression of SFRP1;4.Inhibition of SFRP1 could weaken the effect of down-regulation of miR-586 on U87 cell proliferation and cycle;5.The down-regulation of MiR-586 could inhibit the growth of gliomas;Conclusion:Down-regulation of miR-586 could up-regulate the expression of SFRP1 to inhibit glioma cell proliferation and induce cell cycle arrest.