腺苷酸活化蛋白激酶α在宫颈癌中的表达及其作用机制

Expression and mechanism of adenosine monophosphate-activated protein kinase α in cervical cancer

目的 研究腺苷酸活化蛋白激酶（AMPK）α在宫颈癌中的表达及其作用机制.方法 蛋白质印迹方法检测宫颈癌细胞株HeLa、SiHa细胞中AMPKα1和AMPKα2的表达情况,采用免疫组织化学染色方法检测宫颈癌和正常组织中磷酸化AMPKα （p-AMPKα）的表达情况;采用短干扰RNA（siRNA）降低宫颈癌细胞中AMPKα1和AMPKα2表达水平后,采用蛋白质印迹方法检测相关信号通路、5-溴脱氧尿嘧啶核苷掺入法检测细胞增殖能力.结果 HeLa、SiHa细胞中AMPKα1及AMPKα2均有表达;与正常宫颈组织相比,宫颈癌组织中p-AMPKα表达明显升高[（78.140±2.525）％比（0.209±0.014）％],差异有统计学意义（P＜0.05）.将AMPKα1 siRNA和AMPKα2 siRNA联合转染细胞24 h后,与阴性对照siRNA相比,AMPKα表达水平明显降低,磷酸化雷帕霉素蛋白1（p-mTOR 1）、磷酸化真核生物转录起始因子4E结合蛋白（p-4E-BP1）水平明显升高,细胞增殖能力均明显增强[HeLa细胞：（3.23±0.24）％比（1.50±0.08）％,SiHa细胞：（2.48±0.16）％比（1.11±0.19）％],差异均有统计学意义（均P＜0.05）.结论 宫颈癌细胞中AMPKα活性增强,降低AMPKα水平可促进mTOR1的激活,进而增强肿瘤细胞的增殖.

Objective To investigate the expression and mechanism of adenosine monophosphate-activated protein kinase （AMPK） α in cervical cancer.Methods Western blot was used to detect the expression of AMPKoα1 and AMPKα2 in cervical cancer HeLa and SiHa cell lines;immumohistochemistry was used to detect the expression of phosphorylated-AMPKα （p-AMPKα） in cervical cancer and normal cervical tissue.Short interference RNA （siRNA） targeting to AMPKα1 and AMPKα2 was transfected to HeLa and SiHa cells,then the cell signaling pathway was detected by Western blot and the proliferation of cells was detected by 5-bromodeoxyuridine incorporation method.Results The AMPKα1 and AMPKα2 were both expressed in HeLa and SiHa cells.The expression level of p-AMPKα was significantly higher in cervical cancer tissue than that in normal cervical tissue [（78.140 ± 2.525） % vs （0.209 ± 0.014） %].Compared with negative control siRNA,AMPKα1 siRNA combined with AMPKoα2 siRNA significantly reduced the expression of AMPKα,increased the expressions of phosphorylated-mammalian target of rapamycin 1 （mTOR1） and phosphorylated-eukaryotic initiation factor 4E-binding protein 1,significantly enhanced the cell proliferation ability [HeLa cell line：（3.23 ± 0.24） % vs （1.50 ± 0.08） %,SiHa cell line：（2.48 ± 0.16） % vs （1.11 ± 0.19） %] 24 h after transfection （P 〈 0.05）.Conclusion The activity of AMPKα is significantly increased in cervical cancer;inhibition of AMPKα expression can reduce the proliferation ability of cancer cells through activating mTOR1.