二甲双胍通过AMPK/mTOR信号通路诱导胆囊癌细胞凋亡  

Metformin increases apoptosis of human Gallbladder cancer cells through AMPK/mTOR signaling pathway

作　　者：周汉旭 严华悦 王胜威 刘子祥 周少波 ZHOU Hanxu;YAN Huayue;WANG Shengwei;LIU Zixiang;ZHOU Shaobo(The Second Affiliated Hospital of Bengbu Medical University,Bengbu 233000,China;Anhui Province key Laboratory of Tissue Trasplatation,Bengbu 233000,China)

机构地区：[1]蚌埠医科大学第二附属医院普外科,安徽蚌埠233000 [2]组织移植安徽省重点实验室,安徽蚌埠233000

出　　处：《海南医科大学学报》2025年第6期415-420,共6页Journal of Hainan Medical University

基　　金：安徽省高等学校自然科学研究重点项目(2022AH051476);蚌埠医学院自然科学重点项目(2021byzd197);蚌埠医学院2023年度研究生科研创新计划(Byycx23110)。

摘　　要：目的:本研究旨在探讨二甲双胍通过AMPK/mTOR信号通路对胆囊癌细胞凋亡以及生物学功能产生影响的具体机制。方法:通过使用不同浓度梯度的二甲双胍处理胆囊癌细胞,采用CCK‑8实验法测定二甲双胍对NOZ细胞的抑制效果。实验随后被分为3组:空白对照组、MET组以及MET+BAY‑3827组合。为了评估二甲双胍对NOZ细胞增殖的影响,进行细胞集落形成实验。本研究采用Western blot技术探讨与细胞凋亡相关的蛋白质及AMPK/mTOR信号通路的调控机制。本研究采用了流式细胞术计算胆囊癌细胞中凋亡细胞的比例。细胞划痕实验和Transwell实验被用来评估胆囊癌细胞的迁移能力。结果:二甲双胍可以通过激活AMPK/mTOR信号通路的方式,抑制胆囊癌细胞增殖和迁移,并促进细胞凋亡。CCK‑8和集落形成实验已验证了其对胆囊癌细胞抑制作用,流式细胞术检测到其具有促进胆囊癌细胞凋亡的效果,且使用BAY‑3827抑制该信号通路后,二甲双胍对凋亡相关蛋白的表达趋势的影响有逆转作用,Western blot结果显示,二甲双胍上调凋亡相关蛋白Caspase‑3和Bax以及AMPK/mTOR信号通路相关蛋白AMPK、p‑AMPK的表达,下调抗凋亡蛋白Bcl‑2和AMPK/mTOR信号通路相关蛋白mTOR、p70S6K、4E‑BP1、p‑mTOR、p‑p70S6K、p‑4E‑BP1的表达。结论:二甲双胍可通过上述机制抑制胆囊癌细胞的相关生物学活动,诱导其凋亡。Objective:To investigate the mechanism of metformin on the apoptosis and biological function of gallbladder cancer cells through AMPK/mTOR signaling pathway.Methods:Gallbladder carcinoma cells were treated with different concentrations of metformin,and the inhibitory rate of metformin on NOZ cells was detected using the CCK‑8 assay.The experiments were divided into three groups:blank control group,MET group and MET+BAY‑3827 group.Colony formation assay was performed to detect the effect of metformin on the proliferation of NOZ cells.Western blot was used to detect the expressions of proteins linked to the AMPK/mTOR signaling pathway and proteins related to apoptosis,and flow cytometry was used to estimate the proportion of apoptotic cells.Wound healing assay and transwell assay were used to estimate cell migration efficiency.Results:Metformin inhibited the proliferation and migration of gallbladder carcinoma cells and induced apoptosis via AMPK/mTOR signaling pathway.CCK‑8 assay and colony formation assay showed that metformin could inhibit the proliferation of gallbladder carcinoma cells.Flow cytometry results indicated that metformin increased the ratio of apoptotic cells in gallbladder carcinoma cells,and the trend of metformin's effect on the expression of apoptosis‑related proteins was reversed after inhibiting this signaling pathway with BAY‑3827.Western blot results indicated that metformin upregulated the expression of apoptosis‑related proteins Caspase‑3 and Bax as well as AMPK and p‑AMPK,and downregulated anti‑apoptotic protein Bcl‑2 and AMPK/mTOR signaling pathway‑related proteins mTOR,p70S6K,4E‑BP1,p‑mTOR,p‑p70S6K,and p‑4E‑BP1.Proliferation and migration of human gallbladder carcinoma were inhibited via metformin by activating the AMPK/mTOR signaling pathway,Besides,metformin upregulated the expression of apoptosis‑related proteins Caspase‑3 and Bax and downregulated the expression of anti‑apoptotic protein Bcl‑2,thereby increased the proportion of apoptotic cel

关 键 词：胆囊癌 二甲双胍 信号通路 细胞凋亡 

分 类 号：R735.8[医药卫生—肿瘤]