MicroRNA-653靶向调控OIP5基因介导mTOR信号通路对非小细胞肺癌细胞生物学特性的影响

目的:探讨microRNA-653（miR-653）靶向调控OIP5基因介导mTOR信号通路对非小细胞肺癌（non-small cell lung cancer，NSCLC）细胞生物学特性的影响。方法:选取人正常内皮细胞BEAS-2B和4种NSCLC细胞系（H1650、H1975、A549和H292），分为control组、mimics组、mimics-NC组、inhibitor组和inhibitor-NC组进行瞬时转染，利用荧光定量PCR、蛋白印迹、MTT、划痕实验、Transwell侵袭实验、流式细胞术等方法分析miR-653对NSCLC细胞中OIP5、mTOR信号通路相关基因表达及增殖、迁移、侵袭、凋亡和细胞周期分布的影响。结果:与control组、mimics-NC组和inhibitor-NC组相比，mimics组mTOR信号通路被抑制，细胞增殖、迁移、侵袭能力明显降低，凋亡率明显增加，细胞多停滞于G1期（均P＜0.05）；然而，inhibitor组mTOR信号通路被激活，细胞增殖、迁移、侵袭能力明显升高，而凋亡率明显降低，G1期细胞数量较少（均P＜0.05）。结论:miR-653可通过负调控OIP5基因抑制mTOR通路激活，从而阻止NSCLC的发生与发展。

Effect of microRNA-653 on biological characteristics of human non-small cell lung cancer cells by targeting OIP5 gene and regulating mTOR signaling pathway

Objective:To explore the impact of microRNA-653(miR-653) on proliferation and apoptosis of non-small cell lung cancer(NSCLC) cells by targeting OIP5 gene and regulating mTOR signaling pathway.Methods:Human normal endothelial cell line(BEAS-2B) and four NSCLC cell lines(H1650,H1975,A549 and H292) were assigned into the control,mimics,mimics-negative control(NC),inhibitor and inhibitor-NC groups.Quantitative RT-PCR and Western blot were applied to detect the expressions of miR-653,OIP5 and mTOR.MTT,scratch test,Transwell and flow cytometry were carried out to analyze the effect of miR-653 on cell proliferation,migration,invasion,apoptosis and cycle distribution and regulation of mTOR signaling pathway.Results:Compared with the control and mimics groups,cell proliferation,migration and invasion abilities decreased in the mimics group.Additionally,cell apoptosis rate increased and cells were mostly in G1 phase.miR-653 inhibited the activation of mTOR signaling pathway.Compared with the control and inhibitor-NC group,mTOR signaling pathway was activated and cell proliferation,migration and invasion abilities increased in the inhibitor group,and cell apoptosis rate decreased and cells were less in G1 phase.Conclusion:These results suggest that miR-653 expression was low in NSCLC cells and influenced NSCLC cells by inhibiting OIP5 gene expression and negatively regulating mTOR signaling pathway.