蛋白激酶D2在膀胱癌组织中的表达及其对肿瘤免疫微环境的影响  

Expression of protein kinase D2 in bladder cancer tissue and its effect on tumor immune microenvironment

作　　者：蔡文畅 刘雨齐 王涵 王鹤霖 王振江[3] 肖梓屾 马士远 安丽萍[1] 刘艳波[1,3] CAI Wenchang;LIU Yuqi;WANG Han;WANG Helin;WANG Zhenjiang;XIAO Zishen;MA Shiyuan;AN Liping;LIU Yanbo(National and Local United Engineering R&D Center of Research and Development on Active Peptide of Medicinal Plants and Animals in ChangBai Mountains,Beihua University,Jilin 132013,China;Clinical Laboratory,School Hospital,Beihua University,Jilin 132013,China;Department of Pathophysiology,School of Basic Medical Sciences,Beihua University,Jilin 132013,China)

机构地区：[1]北华大学长白山药用动植物活性多肽研究与开发国家地方联合工程实验室,吉林吉林132013 [2]北华大学校医院检验科,吉林吉林132013 [3]北华大学基础医学院病理生理学教研室,吉林吉林132013

出　　处：《吉林大学学报(医学版)》2025年第2期378-391,共14页Journal of Jilin University:Medicine Edition

基　　金：吉林省科技厅国际合作项目(20210402015GH);吉林省科技厅基础研究项目(YDZJ202201ZYTS194);北华大学研究生创新项目(研创合字2024021和2024066)。

摘　　要：目的:利用生物信息学技术探讨蛋白激酶D2(PRKD2)在膀胱癌(BLCA)组织中的表达及其表达对BLCA患者预后的影响,阐明PRKD2在BLCA发生发展中的作用。方法:从UCSC癌症基因组数据库下载9例正常膀胱样本、19例BLCA癌旁样本和407例BLCA肿瘤样本的数据。采用Mann-Whitney U检验分析PRKD2 mRNA在BLCA肿瘤组织与正常膀胱组织中的表达差异,并利用人类蛋白数据库(HPA)进行蛋白组学的验证。使用R软件的DESeq2包对PRKD2低表达组和PRKD2高表达组BLCA样本进行差异表达基因(DEGs)筛选及鉴定,使用ggplot2包绘制PRKD2的共表达热图,利用基因本体(GO)功能注释和京都基因与基因组百科全书(KEGG)对DEGs进行GO功能注释及KEGG信号通路富集分析,并使用基因富集分析(GSEA)获得DEGs明显富集的基因集。根据PRKD2的表达水平将BLCA样本分为低表达组和高表达组,使用GSVA包分析BLCA患者PRKD2表达与免疫细胞浸润之间相关性。使用survival包和survminer包进一步分析PRKD2与BLCA患者预后的关系。使用cBioPortal数据库对BLCA组织的PRKD2基因突变情况进行分析。收集膀胱炎、膀胱息肉和BLCA组织,使用免疫组织化学染色技术检测BLCA及对照组织中白细胞介素17F(IL-17F)蛋白表达水平。结果:PRKD2在多种恶性肿瘤组织中高表达,BLCA组织中PRKD2 mRNA和蛋白表达水平较正常膀胱组织明显升高(P<0.05)。对PRKD2单基因差异分析共得到1058个DEGs,其中上调DEGs 29个,下调DEGs 1029个。GO功能注释和KEGG信号通路富集分析,DEGs主要富集于涉及感官知觉的化学刺激、卡哈尔体和内肽酶抑制剂活性等生物学过程(BP)以及金黄色葡萄球菌感染通路和年轻人的成熟型糖尿病通路。GSEA分析,DEGs主要富集于Notch信号通路、维甲酸诱导基因I(RIG-I)样受体信号通路、胞质DNA筛选通路、碱基切除修复信号通路、自然杀伤(NK)细胞介导细胞毒性信号通路和T细胞受体信号通路等过程�Objective:To investigate the expression of protein kinase D2(PRKD2)in bladder cancer(BLCA)tissue using bioinformatics analysis method and its effect on the prognosis of BLCA patients,and to clarify the role of PRKD2 in the occurrence and development of BLCA.Methods:The data from 9 normal bladder samples,19 BLCA paracancerous samples,and 407 BLCA tumor samples were downloaded from the UCSC Cancer Genome Database.The Mann-Whitney U test was applied to analyze the difference in expression of PRKD2 mRNA in BLCA tumor and normal bladder tissues,and the Human Protein Atlas(HPA)database was used for proteomic validation.DESeq2 package in R software was applied to screen the differentially expressed genes(DEGs)in BLCA tissue in PRKD2 low-and high-expression groups.The co-expression heatmaps of PRKD2 were plotted using the ggplot2 package,Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)were used for functional annotation analysis and pathway enrichment analysis of DEGs,and Gene Set Enrichment Analysis(GSEA)was used to obtain the gene sets that were significantly enriched for DEGs.The BLCA samples were divided into low-and high-expression groups according to the expression level of PRKD2,and the correlations between PRKD2 expression and immune cell infiltration in the BLCA patients were analyzed with GSVA package.The relationship between PRKD2 and prognosis of BLCA patients was further analyzed using the survival package and the survminer package.The PRKD2 gene mutations in BLCA tissue were analyzed using the cBioPortal database.The cystitis,bladder polyp and BLCA tissues were collected,and the expression levels of interleukin-17F(IL-17F)protein in BLCA and control tissues were detected using immunohistochemical staining technique.Results:PRKD2 was highly expressed in a variety of malignant tumors,and the expression levels of PRKD2 mRNA and protein in BLCA tissue were significantly increased compared with those in normal bladder tissue(P<0.05).Single gene differential analysis of PRKD2 yielded a total of

关 键 词：膀胱肿瘤 蛋白激酶D2 肿瘤免疫微环境 预后标志物 生物信息学分析 

分 类 号：R737[医药卫生—肿瘤]