摘要
目的探究基质Cla蛋白(matrix Cla protein,MGP)在胃癌发病机制中的作用。方法运用免疫组织化学法检测71对胃癌组织及癌旁正常组织中MGP的表达情况,利用基因表达综合(gene expression omnibus,GEO)数据库中2个独立的胃癌数据集进行MGP基因表达分析;并以2株人胃癌细胞系(BGC-823、AGS)为研究对象,应用siRNA质粒敲低MCP,进行MGP敲低后的系列功能实验,如MTS实验、平板集落形成实验、流式细胞术,检测MGP敲低对胃癌细胞的增殖、集落形成能力和凋亡的影响。Western blot检测MCP敲低/过表达对胃癌细胞中凋亡相关蛋白的影响。结果MGP在胃癌组织中的表达明显高于癌旁正常组织(P<0.001),并通过对两个在线数据集的分析得到证实。MGP敲低抑制胃癌细胞BGC-823、AGS的增殖和集落形成(P<0.01);MGP敲低促进胃癌细胞BGC-823.AGS凋亡的发生(P<0.05)。MCP敲低时,胃癌细胞BGC-823、AGS中抗凋亡蛋白BCL-2、cyelinD1表达水平显著下调,而凋亡蛋白CleavedPARP、Caspase3.Bax表达水平显著上调;MGP过表达时,胃癌细胞AGS中BCL-2、cyelin D1抗凋亡蛋白表达水平显著上调,而凋亡标志蛋白Cleaved PARP.Caspase3、Bax表达水平显著下调。结论MCP通过调控胃癌细胞中凋亡相关蛋白的表达,促进胃癌细胞生长并抑制其凋亡,参与胃癌的发生发展。
Objective To explore the role of the matrix Gla protein(MGP)in the pathogenesis of gastric cancer.Methods The expression of MGP in 71 tissues and adjacent normal tissues of gastric cancer was detected by immunohistochemical method and two independent gastric cancer data sets in GEO database were used to analyze MGP gene expression.With two human gastric cancer cell lines(BGC-823,AGS)as the research objects,after using siRNA plasmid to low-expression of MGP,the effects of MGP knockdown on the proliferation,colony formation capacity and apoptosis of gastric cancer cells were detected by a Series of functional experiments about MGP was finished,such as the MTS experiment,the Flat-plate colony formation experiments and the flow cytometry.Western blot tested the effect of MGP low-expression/overexpression on apoptosis-related proteins in gastric cancer cells.Results MGP was expressed significantly higher in gastric cancer tissues than in adjacent normal tissues(P<0.001)and was confirmed by an analysis of two online datasets.MCP knockdown inhibits BGC-823,AGS proliferation and colony formation in gastric cancer cells(P<0.01)and MGP knockdown promotes BGC-823,AGS apoptosis in gastric cancer cells(P<0.05).When MGP was knocked down,the expression level of anti-apoptotic protein in BGC-823 and AGS in gastric cancer cells was significantly lowered,however,the expression level of apoptotic protein Cleaved PARP,Caspase3,Bax was increased signifcantly.With MGP overexpression,BCL-2,cyelin D1 in gastric cancer cell AGS,how-ever the expression level of Cleaved PARP,Caspase3 and Bax was significantly reduced.Conclusion MGP participates in the occurrence and development of gastric cancer by regulating the expression of apoptotic:proteins in gastric cancer cells,promoting the growth and inhibiting the apoptosis of gastric cancer cells.
作者
王觅柱
孟宪梅
党彤
Wang Mizhu;Meng Xianmei;Dang Tong(Department of Gastroe nterology,the Second Afiliated Hospital of Baotou Medical College,Inner Mongolia University of Science and Technology,Baotou 014030,Inner Mongolia,China)
出处
《消化肿瘤杂志(电子版)》
2021年第4期255-262,共8页
Journal of Digestive Oncology(Electronic Version)
基金
内蒙古自治区自然科学基金项目资助(2019MS08054)
内蒙古自治区卫生健康科技计划项目(202201441)。
关键词
基质GLA蛋白
胃癌
凋亡
增殖
Matrix Cla protein
Gastric cancer
Apoptosis
Proliferation