miR-138-5p靶向调控RAB22A表达抑制胶质瘤细胞的增殖、迁移和侵袭

目的:探讨miR-138-5p对胶质瘤细胞增殖、迁移与侵袭的影响以及其作用机制。方法:U251细胞分为miR-NC组(对照组)、miR-mimics组、miR-inhibitor组以及miR-mimics+RAB22A-pcDNA3.1组。qRT-PCR检测miR-138-5p在胶质瘤细胞中的表达量；CCK8实验检测U251细胞的增殖能力；Transwell实验检测U251细胞的迁移和侵袭能力；Targetscan数据库预测miR-138-5p的下游靶基因；运用双荧光素酶报告基因实验验证细胞周期蛋白RAB22A是miR-138-5p的靶基因；RNA结合蛋白免疫沉淀实验进一步验证miR-138-5p与RAB22A之间的相互作用；运用蛋白免疫印迹实验检测组织和细胞中RAB22A的蛋白表达水平。结果:与正常人星形胶质细胞相比较，miR-138-5p 在胶质瘤细胞中表达量降低(P<0.05)；CCK8和Transwell实验结果表明，在U251细胞中，过表达miR-138-5p能显著抑制细胞的增殖、迁移和侵袭能力(P＜0.05)；运用Targetscan数据库预测miR-138-5p下游靶基因为RAB22A，miR-138-5p作用于RAB22A的3' UTR区域，双荧光素酶报告基因实验与蛋白免疫沉淀实验结果证实miR-138-5p与RAB22A之间的相互作用；RAB22A在胶质瘤细胞中明显高表达(P＜0.05)，在U251细胞中过表达miR-138-5p明显抑制RAB22A的表达(P＜0.05)。结论:miR-138-5p在胶质瘤细胞中表达降低，miR-138-5p通过下调RAB22A表达而抑制细胞增殖、迁移和侵袭。

miR-138-5p inhibits proliferation,migration and invasion of glioma cells via targeting RAB22A

Objective:To investigate the effect of miR-138-5p on the proliferation,migration and invading of glioma cells and its mechanism.Methods:U251 cells were divided into miR-NC（control) group,miR-mimics group,miR-inhibitor group and miR-mimics+RAB22A-pcDNA3.1 group.The expression of miR-138-5p in normal human astrocytes and glioma cells was detected by real-time fluorescence quantitative PCR.CCK8 assay was used to explore the proliferation of U251 cells.Transwell assay was used to explore the migration and invasion ability of U251 cells.Targetscan database predicted the downstream target genes of miR-138-5p.Double luciferase reporter gene assay was used to detect that RAB22A was the target gene of miR-138-5p.The interaction between miR-138-5p and RAB22A was further verified by RNA binding protein immunoprecipitation assay.Western blot assay was used to detect RAB22A protein expression in tissues and cells.Results:Compared with normal astrocytes,the expression of miR-138-5p was decreased in glioma cells（P＜0.05).CCK8 and Transwell assay results showed that overexpression of miR-138-5p significantly inhibited the proliferation,migration and invasion of U251 cells（P＜0.05).Targetscan database was used to predict that the downstream target gene of miR-138-5p was RAB22A,and miR-138-5p acted on the 3' UTR region of RAB22A.The interaction between miR-138-5p and RAB22A was verified by dual luciferase reporter gene assay and RNA binding protein immunoprecipitation assay.RAB22A was significantly overexpressed in glioma cells（P＜0.05),and overexpression of miR-138-5p significantly inhibited RAB22A expression in U251 cells（P＜0.05).Conclusion:The expression of miR-138-5p is decreased in glioma cells,and miR-138-5p restrains cell proliferation,migration and invasion by down-regulating RAB22A expression.