摘要
目的肝细胞癌是恶性程度较高的肿瘤之一。我国慢性肝炎患者较多,肝细胞癌发病率较高。本研究旨在筛选、鉴定肝细胞癌相关基因,并对鉴定基因的预后意义作初步分析。方法应用R语言,从GEO数据库中的两个肝细胞癌基因芯片数据GSE112790和GSE101685筛选差异表达基因,并用DAVID数据库对筛选鉴定的差异基因进行GO(Gene Ontology,基因本体论)富集分析、KEGG(Kyoto Encyclopedia of Genes and Genomes京都基因和基因组百科全书)通路分析;采用STRING数据库对差异基因构建PPI(Protein-protein interaction,蛋白互作网络),然后用Cytoscape软件及其插件MOCDE进行可视化和模块筛选。通过GEPIA数据库进行验证,并使用cBioportal数据库汇总关键基因的遗传变异信息。通过RT-PCR(reverse transcription-polymerase chain reaction,实时荧光定量分析)检测正常肝细胞5200和肝癌Hep3B细胞核心关键基因的表达情况。结果从两个数据集共鉴定出51个上调、104个下调的共同差异基因。GO和KEGG通路分析表明上调基因在细胞周期、有丝分裂等显著富集;下调基因在药物代谢和P450有关的各种代谢中显著富集。PPI互作网络筛选出两个功能模块(19个上调基因和8个下调基因),其中ASPM、DLGAP5、PTTG1、CYP2C9、CYP3A4和CYP2E1有6个基因是两个模块的核心关键基因。基因组变异分析中,在ASPM蛋白结构域中发现了一个热点突变Y1725C,RT-PCR检测分析6个核心关键基因的结果与生物信息分析结果一致。结论上调表达的ASPM、DLGAP5、PTTG1基因,以及下调表达的CYP2E1、CYP3A4和CYP2C9基因可能对肝细胞癌诊断及预后具有生物标志物意义;ASPM突变Y1725C高发于肝细胞癌。
Objective Hepatocellular carcinoma is one of the tumors with a high degree of malignancy.There are more patients with chronic hepatitis in China,and the incidence of hepatocellular carcinoma is higher.The purpose of this study is to screen and identify hepatocellular carcinoma related genes,and to make a preliminary analysis of the prognostic significance of identifying genes.Methods Using R language,GSE 112790 and GSE101685 were screened from two hepatocellular carcinoma gene chip data in the GEO database,and the differential gene identification was analyzed by GO,KEGG,The PPI is constructed from the STRING database,and then visualized and filtered by the Cytoscape software and its plug-in MOCDE.Verified through the GEPIA database and using the cBioportal database to summarize information on genetic variation of key genes.RT-PCR(real-time fluorescence quantitative analysis)was used to detect the expression of core key genes in normal hepatocytes 5200 and hepatocellular carcinoma Hep3 B cells.Results A total of 51 upregulated and 104 downgulated common differential genes were identified from the two data sets.The analysis of GO and KEGG pathway showed that the upregulated genes were significantly enriched in cell cycle,mitotic nuclear division,etc.,and the downregulated genes were significantly enriched in drug metabolic process and various metabolisms related to P450 pathway.The PPI network screened out two functional modules(19 up and 8 down),of which ASPM,DLGAP5,PTTG1,CYP2 C9,CYP3 A4 and CYP2 E1 have 6 genes that are the key genes of the two modules.In genomic variation analysis,a hot mutation Y1725 C was found in the ASPM protein domain.The results of RT-qPCR detection and analysis of 6 core key genes are consistent with the results of bioinformatics analysis.Conclusion The study identified 6 genes that may be the key biomarker genes of hepatocellular carcinoma,and the ASPM mutation Y1725 C was high in liver cell carcinoma.
作者
张伟国
郑昊洋
张心怡
蒋飞翔
周兆丽
ZHANG Weiguo;ZHENG Haoyang;ZHANG Xinyi;JIANG Feixiang;ZHOU Zhaoli(Shanghai Key Laboratory of Molecular Imaging,Maying District Central Hospital,Shanghai University of Medicine and Health Sciences,Shanghai 201318,China;School of Health Science and Engineering,University of Shanghai for Science and Technology,Shanghai 200093,China)
出处
《沈阳药科大学学报》
CAS
CSCD
北大核心
2021年第11期1183-1195,共13页
Journal of Shenyang Pharmaceutical University
基金
国家自然科学基金资助项目(81803581)
上海健康医学院2018年协同创新重点专项(SPCI-18-18-003)
上海市分子影像学重点实验室建设项目(18DZ2260400)。
关键词
肝细胞癌
预后
表达谱
差异表达基因
生物信息学分析
hepatocellular carcinoma
prognosis
transcriptomes
differentially expressed genes
bioinformatics
