蛋白酶体抑制剂MG132诱导胃癌细胞AGS凋亡的机制研究  

Mechanism of proteasome inhibitor MG132 induced AGS apoptosis in gastric cancer cells

作　　者：袁婷 韦四喜 夏英 李红羽 杜洪 金泳 黄海 YUAN Ting;WEI Sixi;XIA Ying;LI Hongyu;DU Hong;JIN Yong;HUANG Hai(School of Clinical Laboratory Science,Guizhou Medical University,Guiyang 550004,China;Center for Clinical Laboratories,The Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China;Department of Laboratory Medicine,The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang 550001,China)

机构地区：[1]贵州医科大学医学检验学院,贵州贵阳550004 [2]贵州医科大学附属医院临床检验中心,贵州贵阳550004 [3]贵州中医药大学第一附属医院检验科,贵州贵阳550001

出　　处：《沈阳药科大学学报》2023年第8期1036-1043,共8页Journal of Shenyang Pharmaceutical University

基　　金：贵州省科技创新人才团队项目(2019-5610);临床疾病分子标志物筛选及验证创新团队筑科合同([2019]9-1-5号)。

摘　　要：目的蛋白酶体抑制剂MG132已在几种肿瘤治疗中取得一定疗效,但MG132对胃癌细胞凋亡的作用机制尚不完全清楚。因此,本文主要探讨MG132对胃癌细胞AGS凋亡的影响及其可能机制,旨在为胃癌治疗提供实验基础。方法选择AGS细胞为研究对象,实验分为6个组(MG132浓度分别为0、0.5、1、2.5、5、10μmol·L^(-1)),利用CCK-8和克隆形成实验检测细胞增殖,流式细胞术检测细胞凋亡及活性氧(reactive oxygen species,ROS)水平,全自动生化分析仪检测乳酸脱氢酶(detect lactate dehydrogenase,LDH)水平,免疫沉淀(immunoprecipitation,IP)探讨脑型糖原磷酸化酶(brain-type glycogen phosphorylase,PYGB)、受体相互作用蛋白3(receptor interacting protein 3,RIP3)和受体相互作用蛋白1(receptor interacting protein1,RIP1)的相互作用关系,进一步用Western blot检测PYGB、RIP3和RIP1的表达情况。结果不同浓度MG132处理AGS细胞12、24、36 h后,MG132存在浓度和时间依赖性抑制AGS细胞增殖;MG132处理AGS细胞24 h后,诱导AGS细胞凋亡,促进细胞内LDH释放和ROS水平增加;MG132处理AGS细胞24 h后,细胞内PYGB表达水平降低,RIP3和RIP1表达水平升高。此外,IP实验表明PYGB、RIP3和RIP1之间存在相互作用。结论在胃癌细胞AGS中,MG132诱导的AGS细胞凋亡与PYGB表达水平降低,RIP3和RIP1表达水平升高有关。Objective Proteasome inhibitor MG132 has achieved certain effects in the treatment of several kinds of tumors,but the mechanism of MG132 induced apoptosis in gastric cancer cells is not fully understood.Therefore,in this research we treated AGS cells with different concentrations of MG132,mainly to explore the effect of MG132 on the apoptosis of gastric cancer AGS cells and its possible mechanism.This provided an experimental basis for therapy of gastric cancer.Methods AGS cells were selected as the research object,and the experiment was divided into 6 groups(MG132 concentrations were 0,0.5,1,2.5,5μmol·L^(-1)and 10μmol·L^(-1)),CCK-8 and clone formation experiments were used to detect cell proliferation,flow cytometry were used to detect apoptosis and reactive oxygen species(ROS)levels,automatic biochemical analyzer were used to detect lactate dehydrogenase(LDH)level,Immunoprecipitation(IP)to explore the interaction between brain-type glycogen phosphorylase(PYGB),receptor interacting protein 3(RIP3)and receptor interacting protein 1(RIP1),further used Western blot were used to detect the expression of brain-type glycogen phosphorylase(PYGB),receptor interacting protein 3(RIP3)and receptor interacting protein 1(RIP1).Results After treating AGS cells with different concentrations of MG132 for 12,24 h and 36 h,the proliferation of AGS cells was inhibited in a concentration and time-dependent manner;MG132 induced AGS cell apoptosis,promoted the release of LDH and the increase of ROS levels in the cells for treatment of 24 h;After AGS cells treated with MG132 for 24 h,the expression level of PYGB in the cells decreased,the expression levels of RIP3 and RIP1 in the cells increased.In addition,IP experiments revealed an interaction between PYGB,RIP3,and RIP1.Conclusion In gastric cancer AGS cells,the apoptosis of AGS cells induced by MG132 is related to the decrease of PYGB expression level and the increase of RIP3 and RIP1 expression levels.

关 键 词：胃癌 蛋白酶体抑制剂 脑型糖原磷酸化酶 受体相互作用蛋白3 受体相互作用蛋白1 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]