MDM2的异常表达对胃癌细胞增殖、侵袭、迁移以及对CD4^(+)T和CD8^(+)T细胞免疫浸润的影响  

Effects of Abnormal Expression of MDM2 on Proliferation,Invasion and Migration of Gastric Cancer Cells and Immune Infiltration of CD4^(+)T and CD8^(+)T Cells

作　　者：常攀 魏娜 陈洁[1] CHANG Pan;WEI Na;CHEN Jie(Department of Gastroenterology,the First People′s Hospital of Longquanyi District,Chengdu,Chengdu Sichuan 610100,China)

机构地区：[1]成都市龙泉驿区第一人民医院消化内科,四川成都610100 [2]通用医疗363医院消化内科

出　　处：《联勤军事医学》2024年第7期543-552,共10页Military Medicine of Joint Logistics

基　　金：2019年四川省卫生健康委员会科研课题(19PJ027)。

摘　　要：目的 探讨鼠双微体同源基因2(murine double minute 2,MDM2)在胃癌中的表达,以及MDM2的表达对胃癌细胞增殖、侵袭、迁移以及CD4^(+)T和CD8^(+)T细胞免疫浸润的影响。方法 从基因表达综合(Gene Expression Omnibus,GEO)数据库中提取胃癌芯片数据集GSE62254,统计分析MDM2在胃癌中的表达情况。通过基因表达谱交互分析(Gene Expression Profiling Interactive Analysis,GEPIA)数据库分析MDM2表达对胃癌患者预后的影响。基于单样本基因集富集分析(single-sample gene-set enrichment analysis,ssGSEA)探讨GSE62254中23种免疫细胞的浸润情况。免疫组织化学法检测临床样本组织中MDM2和CD4^(+)T和CD8^(+)T细胞的表达。细胞实验,分为sh组(shRNA-MDM2下调MDM2表达)和sh-NC组(shRNA-MDM2-NC无下调MDM2表达的作用),si组(siRNA-MDM2降低MDM2表达)和si-NC组(siRNA-MDM2-NC无降低MDM2表达的作用),pc组(pcDNA3.1-MDM2过表达MDM2)和pc-NC组(pcDNA3.1-MDM2-NC无过表达MDM2的作用),mimic组(MDM2 mimic上调MDM2表达)和mimic-NC组(MDM2-NC无上调MDM2表达的作用)。动物实验,分为si组(下调MDM2表达)、si-NC组(无下调MDM2表达),mimic组(升高MDM2表达)、mimic-NC组(无升高MDM2表达),每组各5只小鼠。5-乙炔基-2′-脱氧尿苷(5-ethynyl-2′-deoxyuridine,EdU)以及Transwell小室检测各组细胞的增殖、迁移与侵袭性能。动物实验检测各组细胞的体内生长与转移能力;Western blot检测细胞和小鼠瘤体组织中MDM2、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、E-钙黏着蛋白(E-cadherin)和波形蛋白(Vimentin)的表达;同时细胞计数试剂盒(cell counting kit 8,CCK-8)检测小鼠脾组织单细胞悬液中CD8^(+)T细胞的体外杀伤肿瘤细胞的能力。结果 与癌旁正常组织相比,MDM2在胃癌组织的表达明显升高(P<0.05);与MDM2高表达患者相比,MDM2低表达患者的总生存期生存曲线明显升高(P<0.05);与MDM2高表达组相比,MDM2低表达组中CD4^(+)T、CD8^(+)T细胞的浸润Objective To investigate the expression of murine double minute 2(MDM2) in gastric cancer and its effects on the proliferation,invasion,migration and immune infiltration of CD4^(+)T and CD8^(+)T cells.Methods The gastric cancer chip data set GSE62254 was extracted from Gene Expression Omnibus(GEO),and the expression of MDM2 in gastric cancer was statistically analyzed.The effects of MDM2 expression on the prognosis of patients with gastric cancer was analyzed by Gene Expression Profiling Interactive Analysis(GEPIA).The infiltration of 23 kinds of immune cells in GSE62254 was investigated by single-sample gene-set enrichment analysis(ssGSEA).The expressions of MDM2,CD4^(+)T and CD8^(+)T cells in clinical samples were detected by immunohistochemistry.The cell experiments were divided into sh group(siRNA-MDM2 down-regulated MDM2 expression) and si-NC group(siRNA-MDM2-NC not down-regulated MDM2 expression),si group(siRNA-MDM2 depressed MDM2 expression) and si-NC group(siRNA-MDM2-NC not depressed MDM2 expression),pc group(pcDNA3.1-MDM2 overexpressed on MDM2) and pc-NC group(pcDNA3.1-MDM2-NC not overexpressed on MDM2),mimic group(MDM2 mimic up-regulated MDM2 expression) and mimic-NC group(MDM2-NC not up-regulated MDM2 expression).The animal experiments were divided into si group(down-regulated MDM2 expression) and si-NC group(not down-regulated MDM2 expression),mimic group(elevated MDM2 expression) and mimic-NC group(not elevated MDM2 expression),with 5 mice in each.The proliferation,migration and invasion of cells in each group were detected by 5-ethynyl-2′-deoxyuridine(EdU) and Transwell chamber.The growth and metastasis ability of cells in vivo were detected by animal experiments;the expression of MDM2,proliferating cell nuclear antigen(PCNA),E-cadherin and Vimentin in cells and tumor tissues of mice were detected by Western blot;and the in vitro killing ability of CD8^(+) T cells in single cell suspension of mice spleen tissues was detected by cell counting kit 8(CCK-8).Results Compared with adjacent normal tissu

关 键 词：鼠双微体同源基因2 胃癌 增殖 侵袭 迁移 免疫细胞浸润 

分 类 号：R735.2[医药卫生—肿瘤]