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

摘要:

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qRT-PCR和Western blotting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Western blotting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。

Abstract:

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.