二甲双胍通过mTOR途径调节乳腺癌细胞自噬的机制研究  

Metformin may induce autophagy of breast cancer cells by mTOR signaling pathway

作　　者：李少军[1,2] 赵艺涵[3] 高潇 张欣 郭爱宁[1] 刘棣[1] 刁岩[1] 张淑群[1] 闵卫利[1] LI Shaojun;ZHAO Yihan;GAO Xiao;ZHANG Xin;GUO Aining;LIU Di;DIAO Yan;ZHANG Shuqun;MIN Weili(Department of Oncology,the Second Affiliated Hospital of Xi'an Jiaotong University,Shaanxi Xi'an 710004,China;Department of Respiratory and Critical Care Medicine,the First Affiliated Hospital of Xi'an Jiaotong University,Shaanxi Xi'an 710061,China;Hospital of Stomatology,Xi'an Jiaotong University,Shaanxi Xi'an 710004,China)

机构地区：[1]西安交通大学第二附属医院肿瘤科,陕西西安710004 [2]西安交通大学第一附属医院呼吸与危重症医学科,陕西西安710061 [3]西安交通大学口腔医院,陕西西安710004

出　　处：《现代肿瘤医学》2023年第20期3764-3770,共7页Journal of Modern Oncology

基　　金：陕西省国际科技合作重点项目计划项目(编号:2019KW-035);陕西省中西医结合肿瘤防治重点实验室建设项目(编号:2022-ZXY-SYS-002);西安交通大学第二附属医院院基金自由探索[编号:2020YJ(ZYTS)611];西安交通大学第二附属医院新技术(编号:2018-56)。

摘　　要：目的:探讨二甲双胍是否通过哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)途径诱导乳腺癌细胞自噬,抑制肿瘤生长。方法:通过体外培养MCF-7、MDA-MB-231、SK-BR-3三种乳腺癌细胞,给予它们不同浓度的二甲双胍作用24~72 h;通过CCK8、TUNEL、自噬小体实验、电子显微镜、免疫荧光、蛋白印迹分析法等方法检测癌细胞自噬情况。结果:二甲双胍可以抑制MCF-7、MDA-MB-231、SK-BR-3肿瘤细胞的增殖,并且随着二甲双胍的浓度逐渐升高、作用时间逐渐延长,其抑制癌细胞能力增强。二甲双胍可以促进MCF-7肿瘤细胞自噬小体的产生以及细胞的凋亡。MCF-7细胞在给予二甲双胍之后,相比较于对照组,p-AMPK表达增加,p-PI3K、p-AKT以及p-mTOR表达降低;MCF-7细胞在给予二甲双胍+AMPK抑制剂后,相比较于单纯二甲双胍组,p-AMPK表达降低,p-PI3K以及p-AKT表达不变,p-mTOR表达增加;MCF-7细胞在给予二甲双胍+PI3K激动剂后,相比较于单纯二甲双胍组,p-AMPK表达不变,p-PI3K和p-AKT表达增加,p-mTOR表达增加;MCF-7细胞在给予二甲双胍+AKT激动剂后,相比较于单纯二甲双胍组,p-AMPK和p-PI3K表达不变,p-AKT表达增加,p-mTOR表达增加。结论:二甲双胍可能通过激活AMPK/mTOR和抑制PI3K/AKT/mTOR信号通路,诱导MCF-7乳腺癌细胞自噬,发生凋亡,从而抑制癌细胞的增殖。Objective:To investigate whether metformin can induce autophagy of breast cancer cells and inhibit tumor growth through mammalian target of rapamycin(mTOR)pathway.Methods:Three kinds of breast cancer cells,MCF-7,MDA-MB-231 and SK-BR-3,were cultured and treated with different concentrations of metformin for 24~72 hours.The autophagy of cancer cells was detected by CCK8,TUNEL,autophagosome test,electron microscope,immunofluorescence and Western blotting.Results:Metformin could inhibit the proliferation of MCF-7,MDA-MB-231 and SK-BR-3 tumor cells.With the increase of metformin concentration and action time,and cancer cells inhibitory ability was enhanced.Metformin could promote the production of autophagosomes and apoptosis of MCF-7 tumor cells.The expression of p-AMPK was increased but the expression of p-PI3K,p-AKT and p-mTOR was decreased after MCF-7 cells were given metformin,compared with the control group.Compared with the metformin group,the expression of p-AMPK was decreased,the expression of p-PI3K and p-AKT was unchanged,and p-mTOR expression was increased after MCF-7 cells were given metformin+AMPK inhibitor.After the MCF-7 cells were given metformin+PI3K agonist,compared with the metformin group,p-AMPK expression remained unchanged,the expression of p-PI3K and p-AKT was increased,and p-mTOR expression was increased.After MCF-7 cells were given metformin+AKT agonist,compared with metformin group,the expression of p-AMPK and p-PI3K was unchanged,the expression of p-AKT was increased,and the expression of p-mTOR was increased.Conclusion:Metformin may induce autophagy and apoptosis of breast cancer cells MCF-7 by activating AMPK/mTOR and inhibiting PI3K/AKT/mTOR signaling pathway,thus inhibiting the proliferation of cancer cells.

关 键 词：乳腺癌 二甲双胍 自噬 AMPK/mTOR信号通路 PI3K/AKT/mTOR信号通路 

分 类 号：R737.9[医药卫生—肿瘤]