食管鳞癌差异表达基因的筛选及其与免疫细胞浸润和预后的相关性  

Screening of DEGs in esophageal squamous cell carcinoma and correlation analysis with immune cells infiltration and prognosis

作　　者：王高燕 李青园 杨荣芳 王永红 沈素朋[4] WANG Gaoyan;LI Qingyuan;YANG Rongfang;WANG Yonghong;SHEN Supeng(Experimental Center,Hebei University of Chinese Medicine,Shijiazhuang 050200,Hebei,China;Hebei Key Laboratory of Integrated Chinese and Western Medicine for Lung Disease Research,Shijiazhuang 050091,Hebei,China;Center for Disease Control and Prevention of Shijiazhuang,Shijiazhuang 050011,Hebei,China;Hebei Cancer Institute,the Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei,China)

机构地区：[1]河北中医药大学实验中心,河北石家庄050200 [2]河北省中西医结合肺病重点实验室,河北石家庄050091 [3]石家庄市疾控中心,河北石家庄050011 [4]河北医科大学第四医院肿瘤研究所,河北石家庄050011

出　　处：《暨南大学学报（自然科学与医学版）》2024年第4期337-347,共11页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：河北省重点研发计划项目(22377736D);河北省自然科学基金项目(H2022423306,H2022206326);河北省卫健委课题项目(20221489);河北中医学院博士基金项目(BSZ2021025)。

摘　　要：目的:利用GEO数据库筛选食管鳞癌的差异表达基因(DEGs),并进一步分析这些基因与食管鳞癌中免疫细胞浸润之间的相关性,以及对预后的影响。方法:从GEO数据库筛选食管鳞状细胞癌芯片数据集GSE23400,使用R软件中的Limma包来分析DEGs。整合GSE23400芯片中的两平台数据并通过GSVA包的ssGSEA算法计算每个样本免疫细胞浸润程度,并分别计算免疫浸润细胞与DEGs之间的相关性。结合GSE53624数据集分析DEGs对患者生存的影响,分析生存相关基因表达与免疫细胞浸润程度的相关性。使用GSE53622数据集验证食管鳞癌生存相关基因的表达,以及其与患者预后的关系。在38例食管鳞癌和相应癌旁非肿瘤组织中,采用免疫组织化学方法(IHC)检测EHF和CSTB的蛋白表达水平。使用RT-qPCR和Western blot检测EHF和CSTB在食管鳞癌中的mRNA和蛋白表达水平。结果:通过对104例食管鳞癌患者癌组织和正常组织的数据进行分析,共获得38个DEGs。免疫浸润研究发现免疫细胞在癌组织和正常组织中的浸润程度有所不同,38个DEGs与免疫细胞浸润程度具有相关性。在GSE53624数据集中,EHF基因表达与CD4^(+)T细胞和B细胞的浸润程度呈显著正相关,相关系数r分别为0.69和0.67(均P<0.01),CSTB基因表达与CD8^(+)T细胞和自然杀伤T细胞的浸润程度显著正相关,相关系数r分别为0.76和0.62(均P<0.01);食管癌患者中EHF和CSTB基因表达与生存时间显著相关(均P<0.05),EHF和CSTB基因高表达患者的生存时间显著高于低表达患者(中位生存期59.1 vs.24.1个月;63.1 vs.23.9个月)。在GSE53622数据集中,EHF和CSTB基因表达下调(均P<0.01),EHF和CSTB基因高表达的食管鳞癌患者比低表达患者预后好(中位生存期NA vs.29.7个月;NA vs.17.9个月,均P<0.05)。IHC结果显示,EHF和CSTB在食管鳞癌中的蛋白表达水平均低于相应癌旁组织(均P<0.05)。RT-qPCR和Western blot结果显示,EHF和CSTB在食管鳞癌细胞中�Objective:To utilize the GEO database for screening DEGs in tissues of esophageal squamous cell carcinoma.It aims to further analyze the correlation with immune cell infiltration and prognosis.Methods:The dataset GSE23400,containing data on esophageal squamous cell carcinoma,was extracted from the GEO database.DEGs analysis was subsequently performed using the Limma package in the R software.The two-platform data of GSE23400 were integrated and the immune cell infiltration in each sample was calculated by ssGSEA algorithm of GSVA package,and the correlation between immune infiltrated cells and DEGs were calculated.The association between DEGs and the overall survival of patients was investigated using the GSE53624 dataset.Furthermore,the correlation analysis between immune infiltrating cells and genes associated with patient survival was evaluated.Also,the expression levels of the survival-related genes in esophageal squamous cell carcinoma were validated using the GSE53622 dataset,and their correlation with patient prognosis was examined.Immunohistochemistry(IHC)was utilized to detect the protein expression levels of DEGs in 38 cases of esophageal squamous cell carcinoma and the corresponding non-tumor tissues.The mRNA and protein expression levels of EHF and CSTB in esophageal squamous cell carcinoma were detected using RT-qPCR and Western blot.Results:Through the analysis of data from 104 patients with esophageal squamous cell carcinoma,a total of 38 DEGs between cancerous and normal tissues were identified.In immune-infiltration analysis,the immune cell infiltration in cancerous and normal tissue was different,the 38 DEGs were related to the abundance of immune cells infiltration.It was exposed that the expressions of EHF and CSTB genes were significantly correlated with survival time of the patients with esophageal cancer through survival analysis(both P<0.05).In GSE53624 dataset,a significant positive correlation was found between EHF gene expression and CD4^(+)T cells,B cells.The correlation coefficients r

关 键 词：食管鳞癌 EHF基因 CSTB基因 免疫细胞浸润 生存分析 

分 类 号：R735.1[医药卫生—肿瘤]