miR-497-5p靶向叉头蛋白4基因对星形胶质细胞瘤细胞增殖、迁移和侵袭的影响

Effect of miR-497-5p targeting the FOXP4 gene on the proliferation,migration,and invasion of astrocytoma cells

目的探讨miR-497-5p靶向叉头蛋白4(FOXP4)基因对星形胶质细胞瘤细胞增殖、迁移和侵袭的影响。方法采用qRTPCR和Westernblotting检测星形胶质瘤细胞(U87、U251和BT325)和正常星形胶质细胞HA1800中miR-497-5p和FOXP4的表达。采用MTT法和Transwell实验分析过表达miR-497-5p或沉默FOXP4对U251细胞增殖、迁移和侵袭的影响。双荧光素酶报告基因实验和Westernblotting分析miR-497-5p和FOXP4靶向调控的关系。结果3种星形胶质细胞瘤细胞中miR-497-5p的表达水平较HA1800细胞降低(P<0.05),FOXP4的表达水平较HA1800细胞升高(P<0.05)。过表达miR-497-5p后U251细胞增殖、迁移和侵袭(均P<0.05)能力降低。沉默FOXP4后U251细胞增殖、迁移和侵袭(均P<0.05)能力降低。miR-497-5p靶向负性调控FOXP4表达。过表达FOXP4可部分逆转miR-497-5p对U251细胞增殖、迁移和侵袭的抑制作用。结论miR-497-5p靶向下调FOXP4表达抑制星形胶质细胞瘤细胞的增殖、迁移和侵袭。

Objective To investigate the effect of miR-497-5p targeting the FOXP4 gene on the proliferation,migration,and invasion of astrocytoma cells.Methods Quantitative real-time PCR and Western blotting were used to measure the expression of miR-497-5p and FOXP4 in astrocytoma cell lines(U87,U251,and BT325)and normal astrocyte cell line(HA1800).MTT assay and Transwell assay were used to analyze the effect of miR-497-5p overexpression or FOXP4 silencing on the proliferation,migration,and invasion of U251 cells.Dual luciferase reporter gene assay and Western blotting were used to investigate the targeted regulatory relationship between miR-497-5p and FOXP4.Results The expression level of miR-497-5p in the three astrocytoma cell lines was significantly lower than that in HA1800 cells(P<0.05),while the expression level of FOXP4 in the three astrocytoma cell lines was significantly higher than that in HA1800 cells(P<0.05).There were significant reductions in the proliferation,migration,and invasion abilities of U251 cells after miR-497-5p overexpression(all P<0.05),and there were also significant reductions after FOXP4 silencing(all P<0.05).MiR-497-5p targeted and negatively regulated the expression of FOXP4.FOXP4 overexpression partially reversed the inhibitory effect of miR-497-5p on the proliferation,migration,and invasion of U251 cells.Conclusions MiR-497-5p inhibits the proliferation,migration,and invasion of astrocytoma cells by downregulating the expression of FOXP4.