二甲双胍对大鼠催乳素瘤MMQ细胞增殖和凋亡的影响及机制  

Effect of metformin on proliferation and apoptosis of rat prolactinoma MMQ cells and related mechanisms

作　　者：靳凯[1] 阮伦亮 蒲九君 钟艾凌 王福超[1] 谭松[1] 黄华[1] 牟家民 杨刚[1] JIN Kai RUAN Lunliang PU Jiujun ZHONG Ailing WANG Fuchao TAN Song HUANG Hua MOU Jiamin YANG Gang(Department of Neurosurgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China)

机构地区：[1]重庆医科大学附属第一医院神经外科,重庆400016

出　　处：《细胞与分子免疫学杂志》2017年第5期643-648,655,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家临床重点专科建设项目(财社[2011]170号);重庆市科委自然科学基金(cstc2013jcyj A10079)

摘　　要：目的探讨二甲双胍对大鼠催乳素瘤MMQ细胞增殖、细胞周期和凋亡的影响及其机制。方法采用(1.25、2.5、5、10、20)mmol/L二甲双胍处理MMQ细胞48 h,CCK-8法检测细胞增殖能力,流式细胞术检测细胞周期及细胞凋亡,Western blot法检测AMP活化的蛋白激酶α1/2(AMPKα1/2)、磷酸化的AMPKα(p-AMPKα)、哺乳动物雷帕霉素靶蛋白(m TOR)、磷酸化的m TOR(p-m TOR)、胰岛素样生长因子1受体(IGF-1R)、胞外信号调节激酶1/2(ERK1/2)、磷酸化的ERK1/2(p-ERK1/2)、蛋白激酶B(PKB/AKT)、磷酸化的AKT(p-AKT)、P21、细胞周期蛋白依赖激酶4(CDK4)、细胞周期蛋白D1(cyclin D1)、胱天蛋白酶3(caspase-3)、裂解的caspase-3(c-caspase-3)、Bcl-2、Bcl-2相关X蛋白(BAX)的变化。结果二甲双胍处理后,抑制MMQ细胞增殖,使细胞周期阻滞在G0/G1期,诱导细胞凋亡;细胞周期相关蛋白P21增加,CDK4、cyclin D1水平降低;c-caspase-3增加,caspase-3、Bcl-2降低;p-AMPKα水平增加、p-m TOR水平降低;IGF-1R、p-AKT及p-ERK水平降低。结论二甲双胍可能通过激活AMPK/m TOR通路、抑制IGF-1R通路从而抑制MMQ细胞增殖,诱导细胞周期阻滞及细胞凋亡。Objective To investigate the effect of metformin on the cel proliferation,cel cycle and apoptosis of rat prolactinoma MMQ cells in vitro and related molecular mechanisms.Methods The MMQ cells were treated with 1.25,2.5,5,10,20 mmol/L metformin for 48 hours.CCK-8 assay was used to assess the cell proliferation ability;flow cytometry was used to analyze the cell cycle distribution and apoptosis;Western blotting was performed to detect the expressions of AMPKα1/2,p-AMPKα,m TOR,p-m TOR,insulin like growth factor 1 receptor（IGF-1R）,ERK1/2,p-ERK1/2,AKT,p-AKT,P21,CDK4,cyclin D1,caspase-3,cleaved caspase-3（c-caspase-3）,Bcl-2 and BAX.Results Compared with the control group,metformin inhibited cell proliferation,induced cell cycle arrest in the G0/G1 phase and promoted cell apoptosis in MMQ cells.The expressions of P21 and c-caspase-3 increased,meanwhile,the expressions of CDK4,cyclin D1,caspase-3 and Bcl-2decreased by metformin.Besides,the expression of p-AMPKα was elevated,but p-m TOR was reduced.Furthermore,the expressions of IGF-1R,p-AKT and p-ERK descended after metformin treatment.Conclusion Metformin could inhibit cell proliferation,induce cell cycle arrest and apoptosis in MMQ cells by activating AMPK/m TOR signaling pathway and inhibiting IGF-1R signaling pathway.

关 键 词：催乳素瘤 二甲双胍 AMP活化的蛋白激酶(AMPK) 胰岛素样生长因子1受体(IGF-1R) 

分 类 号：R392-33[医药卫生—免疫学] Q279[医药卫生—基础医学]