The Effects Of Androgen Receptor Inhibitor (Flutamide) And AR Gene Silencing On Proliferation And Invasion Of Glioma Cells

Objective To detect the effects of AR inhibitor?flutamide?on the proliferation and invasion of glioma cells.To establish AR gene silencing glioma cell line and explore the effects of AR gene silence in glioma,which lay a foundation for gene targeting therapy about androgen receptor pathway in glioma.Methods?1?Different concentration of flutamine(low concentration1.0×10^-8?medium concentration1.0×10^-7?high concentration1.0×10^-66 mol/L)were added to U251cells.The mRNA and protein expression level of AR was detected by Real Time-PCR and Western Blot.?2?CCK8 detected proliferation of cells;Cell apoptosis was detected by flow cytometry;Effects of invasion by AR inhibitor?flutamide?on U251 cells was evaluated by Transwell assay.?3?The glioma cell lines were established by lentivirus transfection with U251 glioma cell and established stable interference silencing AR gene.The mRNA and protein expression level of AR was detected by Real Time-PCR and Western Blot.?4?CCK8 detected proliferation of cells;Cell apoptosis was detected by flow cytometry;Transwell detects the invasion ability of cells;Xenograft tumors in nude mice were used to detect the effect of AR gene silencing on the ability of glioma cells in vitro.?5?The bax/bcl2 mRNA and protein expression were detected by Real Time-PCR and Western Blot,respectively.Results?1?AR inhibitor?flutamide?significantly inhibited U251 cells proliferation and induced apoptosis in vitro in a dose-defendant manner?P<0.05?.?2?A stable cell line expressing GFP fluorescence was obtained by continuous screening of puromycin for a week.The expression of AR mRNA in U251-AR group was significantly reduced by Real Time-PCR,the difference was statistically significant?P<0.05?.There was no significant difference between the blank and vector-control groups?P>0.05?.?3?CCK8assay showed that the proliferation ability of glioma cells decreased significantly.Flow cytometry revealed an increase in apoptosis.The ability of cell invasion was significantly reduced.In U251-AR group,the tumor volume and weight were significantly samller than that of the blank and vector-control groups.It was suggested that AR gene interference of glioma cells was significantly reduced after the silence?P<0.05?.?4?U251-AR group the expression of anti-apoptotic bcl2 was decreased,while the expression of pro-apoptotic bax was increased compared to the Vector-control and the Blank groups?P<0.05?.Conclusions?1?AR inhibitor?flutamide?significantly inhibited glioma cells proliferation and induced apoptosis in vitro that AR may be a risk factor for the development of gliomas.?2?AR gene silencing glioma cell line was established successfully,which provided a base for further researches about regulation of AR gene in glioma in vitro and in vivo.?3?The proliferation and invasion of U251-AR group were significantly inhibited.And the cell apoptosis of U251-AR group increased significantly.AR gene silencing can inhibit the viability of glioma cells in vitro and in vivo.?4?U251-AR group the expression of anti-apoptotic bcl2 was decreased,while the expression of pro-apoptotic bax was increased.In glioma cells,AR may affect the biological activity of glioma cells by regulating bax/bcl2 apoptotic pathways.