miR-195对肺癌A549细胞生物学行为的调控作用

目的:探讨微小RNA(mi R)-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mi R-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mi R-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(P0.05);此外,细胞中cyclin D1、CDK2、Bcl-2及p-Rb的蛋白水平均显著下降(P0.05)。双萤光素酶报告基因分析显示MYB可能是mi R-195的靶基因,且在过表达mi R-195的A549细胞中回补MYB可部分逆转mi R-195对细胞活力、凋亡及迁移的影响。结论:mi R-195可靶向MYB抑制肺癌A549细胞的生长和迁移,并促进其凋亡。

Regulatory effects of miR-195 on biological behaviors of lung cancer A549 cells

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. METHODS: 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 assay 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 ability 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.