miR-451对食管癌EC9706细胞增殖、凋亡及侵袭能力的影响

Overexpression of miR-451 inhibits cell proliferation and invasion and promotes apoptosis in human esophageal carcinoma cell line EC9706

目的:探讨miR-451对食管癌EC9706细胞增殖、凋亡及侵袭能力的影响.方法:化学合成miR-451mimics,脂质体包裹转染EC9706细胞为miR-451组,同时设立无关序列(Scramble-miR)对照组、脂质体对照组和空白对照组.转染后48h,荧光定量RT-PCR检测miR-451表达量的变化,Westernblot检测Bcl-2、AKT和磷酸化AKT蛋白表达水平,流式细胞仪检测细胞凋亡情况,Transwell侵袭实验检测细胞侵袭能力的改变;MTT法检测转染后l、2、3、4、5、6d各组细胞增殖率.结果:miR-451组的miR-451表达水平显著上调(P<0.01,F=69.26),为空白对照组的15.84倍;miR-451组细胞Bcl-2、AKT和磷酸化AKT蛋白表达均显著下调(P<0.05,F=5.83);miR-451组细胞凋亡率为12.07%±1.12%,与3个对照组比较显著升高(P<0.01,F=26.72);miR-451组平均侵袭细胞数为47.4±7.4,与3个对照组比较显著降低(P<0.01,F=34.55).miR-451组细胞的生长在转染后2d出现显著抑制(P<0.05,F=5.95),并且随时间的延长而日益显著.结论:上调miR-451表达可抑制食管癌EC9706细胞增殖和侵袭,促进细胞凋亡.

AIM：To explore the effect of overexpression of miR-451 on cell proliferation,apoptosis and invasion in human esophageal carcinoma cell line EC9706.METHODS：MiR-451 mimics were constructed and transfected into EC9706 cells using LipofectamineTM2000.EC9706 cells transfected with Scramble-miR,empty liposomes or negativemiR-451 were used as controls.Forty-eight hours after transfection,the expression of miR-451 was detected by RT-PCR,and the expression of Bcl-2,AKT and phosphorylated AKT proteins were detected by Western blot.Cell apoptosis,invasion and proliferation were assessed by flow cytometry,transwell assay and MTT assay,respectively.RESULTS：Compared to control cells,the expression of miR-451 was significantly upregulated（P 0.01,F = 69.26）;the expression of Bcl-2,AKT and phosphorylated AKT proteins was significantly down-regulated（P 0.05,F = 5.83）;cell apoptosis rate significantly increased（P 0.01,F = 26.72）;the average number of cells penetrating Matrigel membrane significantly decreased（P 0.01,F = 34.55）;and cell proliferation was significantly suppressed in a timedependent manner（P 0.05,F = 5.95） in EC9706 cells transfected with miR-451 mimics.CONCLUSION：Over-expression of miR-451 induces apoptosis and suppresses cell proliferation and invasion in human esophageal carcinoma cell line EC9706.