miR-139-5p靶向Notch1抑制肺腺癌细胞增殖、迁移和侵袭

miR-139-5p Targets Notch1 to Inhibit Proliferation,Migration and Invasion of Lung Adenocarcinoma Cells

目的探究miR-139-5p在肺腺癌(lung adenocarcinoma,LUAD)进展中的作用和调控机制。方法实时定量PCR(qRT-PCR)和蛋白质印迹检测LUAD组织和细胞中miR-139-5p和Notch同源物1(Notch1)的表达。经细胞转染后,qRT-PCR和蛋白质印迹验证转染效率。CCK-8、细胞划痕和Transwell实验用于检测LUAD细胞的增殖、迁移和侵袭。双荧光素酶报告基因分析miR-139-5p与Notch1的靶向关系。蛋白质印迹检测PI3K/AKT/mTOR信号通路相关蛋白的表达。结果与正常组织和人正常支气管上皮细胞相比,miR-139-5p在LUAD组织和细胞中显著下调(P<0.01),Notch1显著上调(P<0.01)。与对照组相比,过表达miR-139-5p明显抑制LUAD细胞的增殖、迁移和侵袭,并下调p-PI3K、p-AKT和p-mTOR的表达(P<0.01)。Notch1被确定为miR-139-5p的直接靶标。过表达Notch1减弱了miR-139-5p过表达对LUAD细胞增殖、迁移和侵袭的抑制作用(P<0.05)。与对照组比较,miR-139-5p在体内显著抑制了异形移植瘤的生长(P<0.05)。结论miR-139-5p通过靶向Notch1并失活PI3K/Akt/mTOR途径抑制LUAD细胞的增殖、迁移和侵袭。

Objective To investigate the role and regulatory mechanism of miR-139-5p in the progression of lung adenocarcinoma(LUAD).Methods The expression levels of miR-139-5p and Notch homolog 1(Notch1)in LUAD tissues and cells were detected and the transfection efficiency after cell transfection was verified by Quantitative real-time PCR(qRT-PCR)and Western blotting.The proliferation,migration and invasion of LUAD cells were determined by CCK-8,wound healing and Transwell assay.The targeting relationship of miR-139-5p to Notch1 was analyzed using a dual luciferase reporter gene assay.Western blotting was employed to detect the expression levels of PI3K/AKT/mTOR signaling pathway related proteins.Results miR-139-5p was significantly downregulated(P<0.01)and Notch1 was significantly up-regulated in the LUAD tissues and cells(P<0.01),when compared with those in the normal tissues and human normal bronchial epithelial cells.miR-139-5p overexpression suppressed the proliferation,migration and invasion of LUAD cells,and down-regulated the expression levels of p-PI3K,p-AKT and p-mTOR(P<0.01).Notch1 was identified as a target of miR-139-5p that could directly bind to it.Overexpression of Notch1 attenuated the inhibitory effects of miR-139-5p on proliferation,migration and invasion of LUAD cells(P<0.05).In vivo experiment showed that the overexpression of miR-139-5p inhibited the growth of xenograft tumor in nude mice when compared with that in the control group(P<0.05).Conclusion miR-139-5p inhibits the proliferation,migration and invasion of LUAD cells by targeting Notch1 and inactivation of PI3K/Akt/mTOR pathway.