MARCH1通过PI3K/AKT信号通路对人胃癌细胞迁移和侵袭的促进作用  

Promotion effects of MARCH1 on migration and invasion of human gastric cancer cells through PI3K/AKT signaling pathway

作　　者：王暖 杨丽娟 代娟娟 王爱丽[1] 武艳 刘成霞[1] WANG Nuan;YANG Lijuan;DAI Juanjuan;WANG Aili;WU Yan;LIU Chengxia(Department of Gastroenterology,Affiliated Hospital,Binzhou Medical College,Binzhou 256600,China;Cancer Research Laboratory,Affiliated Hospital,Binzhou Medical College,Binzhou 256600,China)

机构地区：[1]滨州医学院附属医院消化内科,山东滨州256600 [2]滨州医学院附属医院肿瘤研究实验室,山东滨州256600

出　　处：《吉林大学学报（医学版）》2021年第2期352-359,共8页Journal of Jilin University：Medicine Edition

基　　金：山东省教育厅“临床医学+X”学科建设项目(200375);山东省卫健委中医药科技发展计划项目(2019-0517);山东省科技厅重点研发计划项目(2019GSF107099)。

摘　　要：目的:探讨膜相关环状蛋白1(MARCH1)对胃癌细胞迁移和侵袭的影响,并阐明其可能的分子机制。方法:收集胃癌手术切除的20例胃癌组织和癌旁组织,实时荧光定量聚合酶链式反应(RT-qPCR)法和Western blotting法检测不同组织中MARCH1 mRNA和蛋白表达水平。选择人胃黏膜正常上皮细胞GES-1和人胃癌细胞BGC-823、BGC-803、AGS及SGC-7901,RT-qPCR法检测不同细胞中MARCH1 mRNA表达水平。取对数生长期的SGC-7901细胞系分为siNC(转染siNC)、siMARCH1-1组(转染siMARCH1-1)和siMARCH1-2组(转染siMARCH1-2),RT-qPCR法和Western blotting法检测各组细胞中MARCH1 mRNA和蛋白表达水平,CCK-8法检测各组细胞增殖活性,细胞划痕实验检测各组细胞划痕愈合率,Transwell小室实验检测各组细胞侵袭能力,Western blotting法检测各组细胞中磷酸化磷脂酰肌醇3-激酶(p-PI3K)、磷酸化蛋白激酶B(p-AKT)和蛋白激酶B(AKT)蛋白表达水平。结果:与癌旁组织比较,胃癌组织中MARCH1 mRNA和蛋白表达水平明显升高(P<0.05),与胃黏膜正常上皮细胞GES-1比较,人胃癌细胞BGC-823、BGC-803、AGS和SGC-7901中MARCH1 mRNA表达水平明显升高(P<0.05或P<0.01)。与siNC比较,siMARCH1-1组和siMARCH1-2组细胞中MARCH1 mRNA及蛋白表达水平均明显降低(P<0.01);与siNC比较,siMARCH1-1组和siMARCH1-2组细胞增殖活性差异无统计学意义(P>0.05),细胞划痕愈合率和侵袭数均明显降低(P<0.01),细胞中p-PI3K和p-AKT蛋白表达水平明显降低(P<0.01),AKT蛋白表达水平差异无统计学意义(P>0.05)。结论:MARCH1能够促进胃癌细胞的迁移和侵袭,其机制可能与调控PI3K/AKT信号通路有关。Objective:To investigate the effects of membrane associated RING-CH 1(MARCH1)on the migration and invasion of gastric cancer cells,and to clarify its possible molecular mechanism.Methods:A total of 20 cases of gastric cancer tissue and adjacent tissue obtained from gastrectomy were collected,and the expression levels of MARCH1 mRNA and protein in the different tissues were detected by Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)method and Western blotting method.The expression levels of MARCH1 mRNA in the human gastric mucosal normal epithelial cells GES-1 and the human gastric cancer cells BGC-823,BGC-803,AGS and SGC-7901 were detected by RTqPCR method.The SGC-7901 cells in logarithmic growth phase were divided into siNC group(transfected with siNC),siMARCH1-1 group(transfected with siMARCH1-1)and siMARCH1-2 group(transfected with siMARCH1-2).RT-qPCR method and Western blotting method were used to detect the expression levels of MARCH1 mRNA and protein in the cells in various groups.CCK-8 method was used to detect the cell proliferation activities in various groups;cell scratch test was used to detect the scratch healing rates of the cells in various groups;Transwell chamber test was used to detect the cell invasion abilities in various groups.The expression levels of phosphorylated phosphatidylinositol 3-kinase(p-PI3 K),phosphorylated protein kinase B(p-AKT)and protein kinase B(AKT)proteins in various groups were detected by Western blotting method.Results:Compared with the adjacent tissue,the expression levels of MARCH1 mRNA and protein in the gastric cancer tissue were increased(P<0.05).Compared with the human gastric mucosal normal epithelial cells GES-1,the expression levels of MARCH1 mRNA in human gastric cancer cells BGC-823,BGC-803,AGS,and SGC-7901 were all increased(P<0.05 or P<0.01).Compared with siNC group,the expression levels of MARCH1 mRNA and protein in siMARCH1-1 group and siMARCH1-2 group were significantly reduced(P<0.01).Compared with siNC group,the cell proliferation

关 键 词：胃肿瘤 膜相关环状蛋白1 细胞迁移 细胞侵袭 磷脂酰肌醇3-激酶 蛋白激酶B 

分 类 号：R735.2[医药卫生—肿瘤]