山东大学学报 (医学版) ›› 2023, Vol. 61 ›› Issue (9): 101-117.doi: 10.6040/j.issn.1671-7554.0.2023.0468
陈映均,刘同刚
CHEN Yingjun, LIU Tonggang
摘要: 目的 探讨与乙型肝炎病毒(HBV)相关肝细胞癌(HCC)发生发展相关的异常甲基化修饰的差异表达基因及分子机制,以期望用于肝癌的早期诊断。 方法 从公共基因表达数据库(GEO)中下载表达谱芯片GSE121248、GSE107170和DNA甲基化芯片GSE136319,采用R语言筛选HBV相关HCC中癌组织和癌旁组织间差异表达基因(DEGs)和差异甲基化基因(DMGs),并绘制可视化火山图。对甲基化差异表达基因(MDEGs)进行基因本体论(GO)和京都基因和基因组百科全书(KEGG)富集分析,构建蛋白质相互作用(PPI)网络,并使用Cytoscape软件进行分子复合物检测(MCODE)分析和利用cytoHubba插件筛选关键基因。通过癌症基因组图谱(TCGA)数据库验证关键基因的mRNA表达水平,并采用皮尔逊相关系数来评估关键基因在HCC中甲基化和基因表达之间的关系。使用HPA数据库、Cox比例风险回归模型、Kaplan Meier-plotter数据库和ROC分析对关键基因进行蛋白表达、生存分析验证以及预测准确率效能;并分析关键基因的表达与临床指标(肿瘤大小、病理分期)之间的相关性。 结果 从GSE121248和GSE107170数据集中分别筛选出921个和1 172个DEGs,其中下调表达基因分别为570个和714个,上调表达基因分别为351个和458个;DNA甲基化芯片GSE136319数据进行差异分析后,有7 952个高甲基化基因,2 630个低甲基化基因。综合分析DEGs和DMGs,共得到33个低甲基化修饰下表达上调的基因,和158个高甲基化修饰下表达下调的基因。GO富集分析表明,异常甲基化修饰的差异表达基因主要参与羧酸分解代谢、有机酸分解代谢和血红素结合等过程;KEGG通路主要为化学致癌DNA加合物、补体和凝血系统和PPAR信号通路等。STRING和 Cytoscape软件筛选出12个与甲基化表达相关的关键基因,包括FTCD、HRG、C8A、FOXM1、FGA、KLKB1、MBL2、FETUB、TTK、AURKA、PRC1和MAD2L1;经临床样本数据验证,FTCD、HRG、C8A、FOXM1、AURKA、PRC1、TTK和MAD2L1这8个基因在HBV相关HCC患者中差异表达,且与患者预后不良有关;FTCD、HRG、FOXM1、TTK、AURKA、PRC1和MAD2L1的表达水平与肿瘤大小和病理分期相关。 结论 FTCD、HRG、C8A、FOXM1、TTK、AURKA、PRC1和MAD2L1在HBV相关HCC的发病过程中可能起着重要的作用,有可能作为HBV相关HCC的潜在诊断标志物及治疗靶点。
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