miR-195对肺癌A549细胞生物学行为的调控作用

摘要

目的:探讨微小RNA(miR)-195对肺癌细胞株A549生长、凋亡及迁移等生物学行为的影响和其相关作用机制.方法:对体外培养的A549细胞转染miR-195 mimics,分别采用CCK-8法和流式细胞术检测细胞活力、周期分布及凋亡情况;Transwell实验检测细胞的迁移能力;Western blot检测相关调控因子cyclin D1、CDK2、Bcl-2和p-Rb/Rb的蛋白水平;双萤光素酶报告基因分析法预测及验证其可能的靶基因.结果:在A549细胞中过表达miR-195可显著抑制细胞活力并引起细胞周期阻滞,同时细胞迁移率降低,而细胞凋亡率显著上升(P<0.05);此外,细胞中cyclin D1、CDK2、Bcl-2及p-Rb的蛋白水平均显著下降(P<0.05).双萤光素酶报告基因分析显示MYB可能是miR-195的靶基因,且在过表达miR-195的A549细胞中回补MYB可部分逆转miR-195对细胞活力、凋亡及迁移的影响.结论:miR-195可靶向MYB抑制肺癌A549细胞的生长和迁移,并促进其凋亡.%AIM: To investigate the regulatory effects of microRNA(miR)-195 on the biological behaviors, such as viability,apoptosis and migration, of lung cancer A549 cells, and to explore the related mechanisms.METH-ODS:After miR-195 mimics were transfected into the A549 cells,the cell viability, cell cycle distribution and apoptosis were measured by CCK-8 assay and flow cytometry.Transwell assay was used to detect cell migration ability.Furthermore, the protein levels of cyclin D1,CDK2,Bcl-2 and p-Rb/Rb were determined by Western blot.Dual-luciferase reporter as-say was used to screen and identify the possible target genes of miR-195.RESULTS: Over-expression of miR-195 in the A549 cells inhibited the cell viability and induced cell cycle arrest,accompanied with the decrease in the cell migration a-bility and the increase in the apoptotic rate(P<0.05).Furthermore,the protein levels of cyclin D1,CDK2,Bcl-2 and p-Rb were significantly decreased(P<0.05).Dual-luciferase reporter assay demonstrated that MYB was a potential target gene of miR-195.Over-expression of MYB in the A549 cells partially reversed the effects of miR-195 on the cell viability, apoptosis and migration.CONCLUSION: miR-195 inhibits lung cancer A549 cell growth and migration, and promotes cell apoptosis by targeting MYB gene.