基于生物信息学分析YOD1基因在胰腺癌中的作用  

Bioinformatics analysis for the role of YOD1 gene in pancreatic adenocarcinoma

作　　者：席晓明 赵闻侠 李杨 张职铄 邵荣光[1] XI Xiao-ming;ZHAO Wen-xia;LI Yang;ZHANG Zhi-shuo;SHAO Rong-guang(Department of Oncology,Institute of Medicinal Biotechnology,Peking Union Medical College and Chinese Academy of Medical Sciences,Beijing 100050,China;Department of Emergency,Xinhua Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200092,China)

机构地区：[1]中国医学科学院北京协和医学院医药生物技术研究所肿瘤室,北京100050 [2]上海交通大学医学院新华医院急诊科,上海200092

出　　处：《中国医药生物技术》2023年第5期402-414,共13页Chinese Medicinal Biotechnology

基　　金：“重大新药创制”国家科技重大专项(2014ZX09201042);中国医学科学院医学与健康科技创新工程(2021-I2M-1-030)。

摘　　要：目的通过TCGA数据库中胰腺癌的RNAseq,分析YOD1作为胰腺癌预后标志物的潜力及其对肿瘤免疫的影响,为胰腺癌的诊断及预后研究提供参考。方法通过TCGA数据获得RNA数据集;利用R survminer包对数据可视化,survival包分析YOD1对胰腺癌生存期的影响,对于TCGA和GTEx的TPM格式的RNAseq数据,利用ggplot2包可视化数据,pROC包对数据进行分析并绘制ROC曲线;使用linkedomics在线网站对差异基因进行GO和KEGG分析。通过miRWalk、TargetScan、miRDB、miRtarBase共同分析可能调节YOD1表达的miRNA。采用R-project中的GSVA包对数据进行可视化处理,采用免疫浸润算法ssGSEA分析YOD1对免疫细胞的影响;利用TIMER2.0在线数据库分析YOD1对免疫细胞浸润的影响。通过EdU和transwell实验验证YOD1对胰腺腺癌细胞增殖和迁移的影响。通过MTT实验验证YOD1对巨噬细胞增殖的影响。通过Western blot实验验证YOD1对巨噬细胞极化的影响。结果差异分析发现YOD1在胰腺癌中高表达;YOD1高表达患者的总生存期(P=0.028,HR=1.59)和无病生存期(P=0.036,HR=1.52)明显低于YOD1低表达患者;ROC曲线证明YOD1具有成为胰腺癌诊断标志的潜力(AUC=0.953,95%CI=0.931~0.975);通过不同网站预测发现,存在31种miRNAs对YOD1的表达起负性调控作用。KEGG富集分析发现,上述31种miRNAs主要富集在影响癌症进展、p53信号通路和胰腺癌进展;PPI网络分析,预测YOD1主要与VCP、UBC、UBB、UBP4、RPS27A、UBXN6、UFD1L、NPLOC4、FAF2、PLAA发生蛋白-蛋白相互作用;免疫相关分析发现YOD1与Th2免疫细胞在PAAD组织的浸润呈正相关(P<0.05),与TFH细胞的浸润呈负相关(P<0.05);体外实验验证YOD1能够促进胰腺癌细胞迁移、增殖,抑制巨噬细胞的增殖并逆转巨噬细胞由M2型向M1型极化。结论YOD1作为一种去泛素酶,可能通过影响肿瘤微环境来促进胰腺腺癌的发生发展,进而影响胰腺腺癌患者的预后。YOD1有潜力成为胰腺癌诊断�Objective The potential of YOD1 as a prognostic marker of pancreatic adenocarcinoma and its effects on tumor immunity were analyzed by RNAseq of pancreatic adenocarcinoma in the TCGA database to provide the clinical basis for the diagnosis and prognosis of pancreatic adenocarcinoma.Methods RNA data set was available from TCGA data.R survminer package was used for data visualization and survival package was used to analyze the effects of YOD1 on the survival of pancreatic adenocarcinoma.For RNAseq data in TPM format of TCGA and GTEx,ggplot2 package was used to visualize the data and pROC package was utilized to analyze the data and draw the ROC curve.GO and KEGG analyses of differential genes were performed by linkedomics.miRNAs which may regulate the expression of YOD1 were analysed by miRWalk,TargetScan,miRDB and miRtarBase.To explore the effect of YOD1 on immune microenvironment,GSVA package in R-project was used to visualize the data,the effect of YOD1 on immune cells was analyzed by the immunoinfiltration algorithm ssGSEA(built-in algorithm in GSVA package),and the effect of YOD1 on immune cell infiltration was analyzed by TIMER2.0 online database.Finally,the effect of YOD1 on the migration and proliferation of pancreatic adenocarcinoma cells were verified through EdU and transwell experiments,the effect of YOD1 on the proliferation of macrophages was verified through MTT experiments,and the effect of YOD1 on the polarization of macrophages was verified through Western blot experiment.Results The differential analysis showed that YOD1 was highly expressed in pancreatic adenocarcinoma.High expression of YOD1 predicted significantly poorer overall survival(P=0.028,HR=1.59)and relapse-free survival(P=0.036,HR=1.52)for patients.ROC curve demonstrated the potential of YOD1 as a diagnostic marker for pancreatic adenocarcinoma(AUC=0.953,95%CI=0.931-0.975).According to the prediction of different websites,there were 31 kinds of miRNAs that negatively regulated the expression of YOD1.KEGG enrichment analysis showed th

关 键 词：胰腺癌 YOD1 生物信息学 肿瘤分子标志物 

分 类 号：R735.9[医药卫生—肿瘤]