过表达miR-34a对肾癌细胞株769-P生物学行为的影响

The effect of overexpression of miR-34a on the biological behavior of renal cancer cell line 769-P

目的探讨microRNA-34a(miR-34a)外源性表达对人肾癌细胞769-P细胞生物学行为的影响。方法建立稳定表达的空载体(NC)与miR-34a细胞株,分为空白对照组、NC组、miR-34a组,qRT-PCR检测miR-34a在各肾癌细胞系中的表达水平,细胞增殖实验、Transwell迁移实验、流式细胞周期检测观测miR-34a高表达后769-P细胞的增殖、细胞周期及迁移能力的变化情况。结果与空白对照组相比,miR-34a组细胞增殖能力降低,在48、72h增殖明显受到抑制,穿膜细胞数目明显减少(P<0.025)。空白对照组、NC组、miR-34a组迁出细胞数分别为:(382.18±37.84)、(398.57±24.09)、(103.73±17.36)个,差异具有统计学意义(P<0.05)。空白对照组、NC组、miR-34a组S期的细胞比例分别为(37.31±2.12)%、(35.96±1.78)%、(57.24±2.67)%,miR-34a组S期的细胞比例比另两组显著升高(P<0.025)。结论miR-34a在生物学行为上可降低肾癌细胞的恶性进展,有望在未来肾癌靶向治疗中发挥抑癌基因作用。

Objective To investigate the effect of exogenous expression of microRNA-34a on the biological behavior of human renal cancer cell line 769-P.Methods The stable expression of 769-P/NC and 769-P/miR-34a cell lines were established by infecting 769-P with miR-34a expression lentivirus vector and control.They were divided into three groups:769-P,769-P/NC and 769-P/miR-34a.The expression of miR-34a in renal cancer cell lines was detected by real-time quantitative reverse transcriptase polymerase reaction(qRT-PCR).The proliferation,cell cycle and migration ability of 769-P cells were detected by cell proliferation,transwell migration and flow cytometry.Results Compared with the control group,the overexpression of miR-34a decreased the proliferation of 769-P/miR-34a cells at 48h and 72h(P<0.05),and the number of transmembrane cells decreased significantly.The cell counts of 769-P,769-P/NC and 769-P/miR-34a were 382.18±37.84,398.57±24.09 and 103.73±17.36(P<0.01).The percentage of S-phase cells in 769-P/miR-34a group was significantly higher than that in the other two groups,which were(37.31±2.12)%,(35.96±1.78)%and(57.24±2.67)%,respectively.Conclusion MiR-34a is expected to play a role of tumor suppressor gene in targeted therapy of renal cancer in the future.