肝癌差异表达基因的生物信息学分析及CSMD2在肝癌中的作用初探

发布时间：2018-05-25 22:46

本文选题：肝癌 + 差异表达基因　；参考：《重庆医科大学》2017年硕士论文

【摘要】：第一部分肝癌差异表达基因的生物信息学分析目的:通过生物信息学方法分析HCC(Hepatocellular Carcinoma)肝癌相关差异表达基因,筛选HCC判断预后的分子标志物和免疫治疗的潜在分子靶点。方法:应用公共基因表达数据库(Gene Expression Omnibus,GEO)下载肝癌相关芯片数据,采用JMP软件进行GSE数据集相关性分析,使用R语言中的Limma程序筛选差异表达基因,并对差异表达基因进行GO(Gene Ontology)富集分析、KEGG(Kyoto Encyclopedia of Genes and Genomes)通路分析及蛋白互作(protein-protein interaction,PPI)调控网络构建。同时联合TCGA数据库中其他肿瘤RNA-Seq转录组数据进行HCC特异性表达分析,进一步筛选HCC特异差异表达基因。结果:MBL2、SDS、SLCO1B3、TDO2、SAA4、SPP2在HCC中特异性表达下调,GPC3基因在肝癌中表达上调,只有SPP2的表达与患者的生存期正相关。结论:GPC3基因可作为HCC免疫治疗的潜在分子靶点,SPP2有望成为判断肝癌患者预后的标志物。第二部分CSMD2在肝癌中的作用初探目的:对CSMD2基因进行系统进化分析和功能注释,检测其在HCC组织和癌旁组织中的表达,并探讨CSMD2在HCC中的作用。方法:通过NCBI数据库检索CSMD2序列,采用Tree Fam网站构建CSMD2系统进化树,对CSMD2进行功能注释,q RT-PCR技术及免疫组化方法分别检测CSMD2在HCC组织的表达,分析CSMD2在不同临床病理特征HCC病人的表达情况。结果:在RNA水平上,CSMD2在癌旁组织中的表达高。免疫组化方法检测,CSMD2在癌旁组织中阳性表达率为52.0%,在HCC组织的阳性表达率为84.0%。CSMD2在肝癌组织内的表达与患者的性别、年龄、术前AFP、巴塞罗那分期、病理分级、HBs Ag阳性与否,肿瘤的大小及数目等无相关性,CSMD2在HCC患者的表达缺失与患者的淋巴结转移正相关(p0.05)。结论:CSMD2的表达缺失可能导致HCC的转移。
[Abstract]:Part I Bioinformatics Analysis of differentially expressed genes in Hepatocellular carcinoma objective: to analyze the differentially expressed genes related to HCC(Hepatocellular carcinoma by bioinformatics, and to screen the molecular markers for predicting the prognosis of HCC and the potential molecular targets for immunotherapy. Methods: using common gene expression database (Gene Expression Omnibus GE) to download liver cancer related chip data, JMP software was used to analyze the correlation of GSE data sets, and Limma program in R language was used to screen differentially expressed genes. The differentially expressed genes were analyzed by GO(Gene Ontology) enrichment analysis and the construction of protein interaction protein-protein interaction (PPI) regulatory network. At the same time, HCC specific expression was analyzed with other tumor RNA-Seq transcriptional data in TCGA database to further screen HCC specific differentially expressed genes. Results the specific expression of TDO2SAA4SPP2 in HCC was down-regulated. The expression of GPC3 gene was up-regulated in HCC. Only the expression of SPP2 was positively correlated with the survival time of the patients. Conclusion the molecular target of HCC immunotherapy, SPP2, may be a potential marker for predicting the prognosis of HCC patients. Part two the role of CSMD2 in hepatocellular carcinoma objective: to investigate the expression of CSMD2 gene in HCC and adjacent tissues, and to explore the role of CSMD2 in HCC by phylogenetic analysis and functional annotation. Methods: CSMD2 sequences were retrieved from NCBI database, and CSMD2 phylogenetic tree was constructed by Tree Fam website. The expression of CSMD2 in HCC tissues was detected by using q-RT-PCR technique and immunohistochemical method, respectively. To analyze the expression of CSMD2 in HCC patients with different clinicopathological features. Results: the expression of CSMD2 in paracancerous tissues was high at RNA level. Immunohistochemical method was used to detect the positive expression rate of CSMD2 in paracancerous tissues (52.0%), and the positive expression rate of 84.0%.CSMD2 in HCC tissues was the same as sex, age, preoperative AFP, Barcelona stage, pathological grade and HBs Ag positive or not. There was no correlation between the size and number of tumors and the loss of CSMD2 expression in patients with HCC. There was a positive correlation between the loss of CSMD2 expression and lymph node metastasis (p0.05). Conclusion the loss of the expression of HCC might lead to the metastasis of HCC.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.7

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