蛋白酶体抑制剂MG132对人胃癌细胞MGC-803自噬、凋亡和迁移的影响  

Effects of proteasome inhibitor MG132 on autophagy,apoptosis and metastasis in human gastric cancer cells MGC-803

作　　者：杨奇 许跃 YANG Qi;XU Yue(Department of Pathology,Zhenjiang Hospital of Chinese Traditional and Western Medicine,Zhenjiang 212002,China;Department of Pathology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]镇江市中西医结合医院病理科,江苏镇江212002 [2]郑州大学第一附属医院病理科,河南郑州450052

出　　处：《肿瘤基础与临床》2024年第2期125-130,共6页journal of basic and clinical oncology

基　　金：河南省科技攻关计划项目(222102310099)。

摘　　要：目的探究蛋白酶体抑制剂MG132对人胃癌细胞MGC-803自噬、凋亡和迁移的影响,分析其对上皮间质转化(EMT)相关蛋白表达的影响。方法应用0.25μg/mL、0.50μg/mL、1.00μg/mL和2.00μg/mL的MG132处理MGC-803细胞后,用激光共聚焦显微镜检测自噬小体数目的变化以分析其对自噬的影响,通过流式细胞仪检测细胞凋亡比例的变化,并通过Transwell细胞迁移实验检测MG132对细胞迁移能力的影响。通过免疫印迹实验检测细胞凋亡相关蛋白和EMT相关蛋白表达的变化。结果在MGC-803细胞中,与空白对照组比较,随着MG132药物浓度的升高,MG132对MGC-803细胞具有毒性作用;1.00μg/mL的MG132明显增加MGC-803细胞的自噬小体数目(1.00μg/mL:t=9.459,P=0.011);随着MG132浓度的升高,细胞凋亡比例明显增多(0.25μg/mL:t=5.149,P=0.036;0.50μg/mL:t=7.342,P=0.018;1.00μg/mL:t=15.340,P=0.004);Western blotting发现MG132处理MGC-803细胞后,随着药物浓度的增加,BAX表达明显增加(0.25μg/mL:t=4.646,P=0.043;0.50μg/mL:t=4.610,P=0.044;1.00μg/mL:t=10.760,P=0.009),Bcl-2表达明显降低(0.25μg/mL:t=22.850,P=0.002;0.50μg/mL:t=26.780,P=0.001;1.00μg/mL:t=29.890,P=0.001);随着MG132浓度的升高,迁移细胞数目明显减少(0.25μg/mL:t=16.730,P=0.004;0.50μg/mL:t=27.340,P=0.001;1.00μg/mL:t=32.800,P<0.001);0.50μg/mL浓度MG132改变EMT相关蛋白表达水平,E-钙黏蛋白表达明显升高(0.50μg/mL:t=4.405,P=0.048;1.00μg/mL:t=12.170,P=0.007),N-钙黏蛋白(0.50μg/mL:t=5.163,P=0.036;1.00μg/mL:t=6.811,P=0.021)和波形蛋白(0.50μg/mL:t=5.628,P=0.030;1.00μg/mL:t=12.670,P=0.006)表达明显降低。结论MG132处理人胃癌细胞MGC-803能促进细胞自噬和凋亡,并通过EMT进程抑制细胞迁移。Objective To investigate the effects of proteasome inhibitor MG132 on the autophagy,apoptosis and metastasis of human gastric cancer MGC-803 cells,and analyze the effects of proteasome inhibitor MG132 on the expression of proteins related to epithelial mesenchymal transformation(EMT).Methods MGC-803 cells were treated with different concentrations(0.25μg/mL,0.50μg/mL,1.00μg/mL and 2.00μg/mL)of MG132,the number of autophagosomes were detected by laser confocal microscopy,the proportion of cell apoptosis was detected by flow cytometry and the effect on cell metastasis was detected by Transwell assay.The expression of apoptosis-related proteins and EMT-related proteins were detected by Western blotting assay.Results Compared with the blank control group,MG132 had toxic effects on MGC-803 cells with the increase of MG132 concentration.MG132 at 1.00μg/mL significantly increased the number of cell autophagosomes in MGC-803 cells(1.00μg/mL:t=9.459,P=0.011).With the increase of MG132 concentration,the apoptosis percentage increased significantly(0.25μg/mL:t=5.149,P=0.036;0.50μg/mL:t=7.342,P=0.018;1.00μg/mL:t=15.340,P=0.004).Western blotting suggested that after MG132 treatment of MGC-803 cells,the expression of BAX expression gradually increased(0.25μg/mL:t=4.646,P=0.043;0.50μg/mL:t=4.610,P=0.044;1.00μg/mL:t=10.760,P=0.009)and Bcl-2 protein expression gradually decreased(0.25μg/mL:t=22.850,P=0.002;0.50μg/mL:t=26.780,P=0.001;1.00μg/mL:t=29.890,P=0.001).In addition,MG132 inhibited the number of migrating MGC-803 cells(0.25μg/mL:t=16.730,P=0.004;0.50μg/mL:t=27.340,P=0.001;1.00μg/mL:t=32.800,P<0.001);furthermore,0.50μg/mL MG132 changed the expression level of EMT-related proteins,and the expression of E-cadherin significantly increased(0.50μg/mL:t=4.405,P=0.048;1.00μg/mL:t=12.170,P=0.007),N-cadherin(0.50μg/mL:t=5.163,P=0.036;1.00μg/mL:t=6.811,P=0.021)and Vimentin(0.50μg/mL:t=5.628,P=0.030;1.00μg/mL:t=12.670,P=0.006)expression gradually decreased.Conclusion MG132 could promote autophagy and apoptosis

关 键 词：蛋白酶体抑制剂 胃癌 自噬 凋亡 迁移 

分 类 号：R735.2[医药卫生—肿瘤]