膜穿孔蛋白E在上皮性卵巢癌...织中的表达水平及其临床意义_陈猛猛.pdf

第 49 卷 第 2 期2023年 3 月吉林大学学报（医学版）Journal of Jilin University（Medicine Edition）Vol.49 No.2Mar.2023DOI：10.13481/j.1671587X.20230220膜穿孔蛋白 E在上皮性卵巢癌组织中的表达水平及其临床意义陈猛猛1,陈俊宇2,高燕1,房正轩3,赵淑华2,郑晶莹2,刘淑香1（1.吉林大学护理学院康复治疗学教研室，吉林 长春 130021；2.吉林大学第二医院妇产科，吉林 长春 130021；3.延边大学药学院药剂学教研室，吉林 延吉 133002）摘要 目的目的：探讨膜穿孔蛋白 E（GSDME）在上皮性卵巢癌（EOC）组织中的表达水平，阐明GSDME 对 EOC 发 生 发 展 的 影 响。方 法方 法：基 于 肿 瘤 基 因 组 图 谱（TCGA）和 基 因 型-组 织 表 达（GTEx）数据库，利用 R 语言分析正常组织和 EOC 组织中 GSDME mRNA 表达水平及其表达水平与EOC 发 生 的 关 系。利 用 差 异 性 分 析、基 因 本 体 论（GO）分 析、京 都 基 因 与 基 因 组 百 科 全 书（KEGG）通路富集分析、基因集富集分析（GSEA）、蛋白-蛋白互作（PPI）网络、HUB基因互作网络和免疫浸润分析探讨与 GSDME 相关联的基因、信号通路及其对 EOC 发生发展的影响。收集 40例EOC 患者组织病理切片，按病理分级分为低级别（中分化及高分化，24 例）组和高级别（低分化，16例）组，按照对顺铂（DDP）的敏感程度分为 DDP耐药组和 DDP敏感组。另收集正常卵巢、良性卵巢瘤和交界性卵巢癌组织切片各 5例作为对照，利用免疫组织化学（IHC）法检测所有组织切片中GSDME 的表达强度。取 10对新鲜的卵巢癌组织和癌旁组织，采用 Western blotting法检测 2组样本中GSDME 蛋白表达水平。体外培养 Skov3 和 Skov3/DDP 细胞，采用 Western blotting 法检测细胞中GSDME 蛋白表达水平，分析其与卵巢癌耐药的关系。结果结果：TCGA 和 GTEx数据库中非配对样本分析，与正常卵巢组织比较，EOC 组织中 GSDME mRNA 表达水平明显降低（0.05），且高级别组EOC 组织中 GSDME mRNA 表达水平低于低级别组（1，0.05。功能富集及免疫浸润分析，GSDME 高表达可以上调循环免疫球蛋白介导的体液免疫反应通路、白细胞迁移通路和免疫球蛋白复合体通路的活性，促进肿瘤微环境中免疫细胞的浸润。IHC 染色，GSDME 与 EOC 的病理组织分级及耐药有关，其中 DDP敏感组 EOC 组织中 GSDME 表达强度明显高于 DDP耐药组（0.05）。Western blotting法检测，与癌旁组织比较，EOC 组织中 GSDME 蛋白表达水平明显降低（0.05）；与 Skov3细胞比较，Skov3/DDP 细胞中 GSDME 蛋白表达水平明显降低（0.05）。结论结论：GSDME 与 EOC 的恶性程度和耐药表现有密切关联。GSDME 低表达是导致肿瘤免疫浸润下降、EOC恶化及耐药的一个关键因素。GSDME可能成为治疗 EOC的潜在分子靶点。关键词 膜穿孔蛋白 E；上皮性卵巢肿瘤；基因表达；生物信息学中图分类号 R737.31 文献标志码 AExpression level of gasdermin E in epithelial ovarian cancer tissue and its clinical significanceCHEN Mengmeng1,CHEN Junyu2,GAO Yan1,FANG Zhengxuan3,ZHAO Shuhua2,ZHENG Jingying2,LIU Shuxiang1文章编号 1671587X(2023)02044012收稿日期 20220807基金项目 吉林省科技厅自然科学基金项目（20200201589JC）作者简介 陈猛猛（1995），男，河南省信阳市人，在读硕士研究生，主要从事肿瘤康复方面的研究。通信作者 郑晶莹，医师（E-mail：zheng_）；刘淑香，副教授（E-mail：）440陈猛猛，等.膜穿孔蛋白 E在上皮性卵巢癌组织中的表达水平及其临床意义（1.Department of Rehabilitation Therapy，School of Nursing，Jilin University，Changchun 130021，China；2.Department of Obstetrics and Gynecology，Second Hospital，Jilin University，Changchun 130021，China；3.Department of Phamaceutics，School of Pharmacy，Yanbian University，Yanji 133002，China）ABSTRACT Objective：To discuss the expression level of gasdermin E（GSDME）in the epithelial ovarian cancer（EOC）tissue，and to clarify the effect of GSDME on the occurrence and development of EOC.Methods：The expressions of GSDME mRNA in normal tissue and EOC tissue was detected by R language based on the Cancer Genome Atlas（TCGA）and Genotype-Tissue Expression（GTEx）Databases；the correlation between the expression level of GSDME and the occurrence of EOC was analyzed.The difference analysis，Gene Ontology（GO）analysis，Kyoto Encyclopedia of Genes and Genomes（KEGG）pathway richment analysis，Gene Set Enrichment analysis（GSEA），protein-protein interaction（PPI）network，HUB gene interaction network，and immune infiltration analysis were used to discuss the genes and signaling pathways associated with GSDME and their effects on the occurrence and development of EOC.The pathological tissue sections of 40 EOC patients were collected and divided into low-grade group（well-differentiated and moderately-differentiated，n=24）and high-grade group（poorly differentiated，n=16）based on the pathological grades；according to the sensitivity to cisplatin（DDP），the pathological tissue sections were divided into DDP resistant group and DDP sensitive group.In addition，5 tissue sections of normal ovary，benign ovarian tumor，and 5 tissue sections of borderline ovarian carcinoma were collected as controls.The expression intensities of GSDME in all the tissue sections were detected by immunohistochemistry（IHC）method.Besides，10 pairs of fresh ovarian cancer and paracancerous tissues were selected，and the expression levels of GSDME protein in the samples in two groups were detected by Western blotting method.The ovarian cancer Skov3 cells and Skov3/DDP cells were cultured in vitro，and the expression levels of GSDME protein in the cells were detected by Western blotting method and the relationship between the expression level and DDP-resistance of ovarian cancer was analyzed.Results：The analysis results of unpaired samples in the TCGA and GTEx Databases showed that compared with normal ovarian tissue，the expression level of GSDME mRNA in EOC tissue was significantly decreased（P0.05），and the expression of GSDME mRNA in EOC tissue in high-grade group was higher than that in low-grade group（P1，P0.05）.The functional enrichment and immune infiltration analysis results showed that high expression of GSDME could upregulate the activities of humoral immunie response pathways，leukocyte migration pathway and immunoglobulin complex pathway mediated by circulating immunoglobulin and promote the infiltration of immune cells in tumor microenvironment.The IHC staining results showed that GSDME was related to the pathological grade and drug resistance of EOC；the expression intensity of GSDME in EOC tisssue in DDP sensitive group was higher than that in DDP resistant group（P0.05）.The Western blotting