MicroRNA-490-5p靶向CDK1通过ERK信号通路参与胃癌细胞周期调控

摘要

目的:探讨miR-490-5p对胃癌细胞增殖与周期的调控作用和分子机制,以期为临床治疗胃癌提供新的有效靶点.方法:收集30例胃癌患者的胃癌组织及相应的癌旁组织标本;采用Real-time PCR法检测miR-490-5p和CDK1的表达水平;分析细胞周期相关蛋白 CDK1 与 miR-490-5p 的靶向关系.MTT法和流式细胞仪检测转染后胃癌细胞生长以及细胞周期情况.结果:与癌旁组织相比,胃癌组织中miR-490-5p的表达显著下调(P<0.001);转染 miR-490-5p mimics 的细胞中miR-490-5p的表达显著上调,而miR-490-5p inhibitors中miR-490-5p显著下降(均P<0.001);CDK1 是 miR-490-5p 的靶基因;下调miR-490-5p、上调CDK1能促进胃癌细胞的增殖能力及G1/S期的转化(均P<0.001).结论:通过上调miR-490-5p 可以抑制胃癌细胞的恶性增殖,减少CDK1表达,抑制 ERK信号途径,降低了G1/S期的转化速率,从而为胃癌诊断及治疗提供新靶标.%Objective:To investigate the effects of microRNA-490-5p(miR-490-5p)on the cell proliferation and cell cycle progression in gastric cancer(GC).Methods:Thirty GC tissues and relative adjacent normal tissues were collected.Quantitative real-time polymerase chain reaction(qRT-PCR)was performed to detect the expression of miR-490-5p and cyclin dependent kinases(CDK).The target relationship of miR-490-5p and CDK1 was analyzed.Cell growth and cell cycle were detected using MTT assay and flow cytometry,respectively.Results:miR-490-5p was significantly down-regulated in GC tissues compared to that in the adjacent normal tissues(P<0.001).miR-490-5p was highly expressed in the cells transfected with miR-490-5p mimics but significantly down-regulated in those transfected with the miR-490-5p inhibitors(both P<0.001).miR-490-5p targeted CDK1.Down-regulating miR-490-5p and up-regulating CDK1 expression promoted the proliferation and G1/S transition of GC cells by targeting CDK1(all P<0.001).Conclusion:The proliferation of GC cells could be inhibited by up-regulating miR-490-5p.Over-expressed miR-490-5p could decrease the expression of CDK1 and inhibit ERK signaling pathway,which suppress the GC cell proliferation and G1/S transition.