Identification of potential miRNA-mRNA regulatory network contributing to pathogenesis of HBV-related HCC
Identification of potential miRNA-mRNA regulatory network contributing to pathogenesis of HBV-related HCC
复制标题
鉴定有助于 HBV 相关 HCC 发病机制的潜在 miRNA-mRNA 调控网络
DOI：
10.1186/s12967-018-1761-7
复制
发表时间：
2019-01-03
影响因子：
7.4
通讯作者：
Fan, Weimin
中科院分区：
文献类型：
--
作者：
Lou, Weiyang;Liu, Jingxing;Fan, Weimin
背景B型肝炎病毒（Hepatitis B virus，HBV）是肝细胞癌（Hepatocellular carcinoma，HCC）的主要危险因素之一。越来越多的证据表明microRNA（miRNA）-mRNA轴参与了HCC的发生。然而，在HBV相关的HCC中仍然缺乏一个全面的miRNA-mRNA调控网络。本研究的目的是确定潜在的miRNA-mRNA调控通路有助于HBV相关的HCC的发病机制。GEO 2 R和R-limma'进行差异表达分析。将两组分析集中出现的共同miRNA筛选为潜在的差异表达miRNA（DE-miRNAs）。使用Kaplan-Meier RT-PCR数据库进一步评估筛选的DE-miRNAs在HCC中的预后作用。通过miRNet预测DE-miRNAs的靶基因。然后，通过STRING数据库为这些靶点建立蛋白质-蛋白质相互作用（PPI）网络，然后通过Celloscape识别网络中的枢纽基因。通过大卫数据库对靶基因进行功能注释和途径富集分析。选取3条HBV相关HCC的富集通路进行进一步分析，筛选出3条通路中常见的潜在靶基因。利用Cytoscape构建miRNA-hub基因网络。采用qRT-PCR方法进一步检测HBV相关肝癌组织中DE-miRNAs的表达及其与靶基因的相关性。7个DE-miRNAs中有5个表达上调，5个表达下调，提示其在HCC中具有重要的预后作用。分别预测了2312和1175个靶基因的上调和下调DE-miRNA。TP 53被鉴定为PPI网络中的枢纽基因。途径富集分析表明，这些预测的靶点与B型肝炎、癌症中的途径、癌症中的microRNA和病毒致癌有关。对这些途径的进一步分析分别筛选了20个和16个上调和下调DE-miRNA的靶基因。通过检测36个靶基因的表达，筛选出6个候选靶基因。最后，根据qRT-PCR结果和表达相关性分析，建立了一个潜在的miRNA-mRNA调控网络。结论本研究初步确定了miRNA-mRNA调控通路，为HBV相关性肝癌的发病机制和有效治疗策略提供了新的思路。
BackgroundHepatitis B virus (HBV) is one of the major risk factors of hepatocellular carcinoma (HCC). Increasing evidence indicates that microRNA (miRNA)-mRNA axis is involved in HCC. However, a comprehensive miRNA-mRNA regulatory network in HBV-related HCC is still absent. This study aims to identify potential miRNA-mRNA regulatory pathways contributing to pathogenesis of HBV-related HCC.MethodsMicroarray GSE69580 was downloaded from Gene Expression Omnibus (GEO) database. GEO2R and R-limma' were used to conduct differential expression analysis. The common miRNAs appeared in the two analytic sets were screened as potential differentially expressed miRNAs (DE-miRNAs). The prognostic roles of screened DE-miRNAs in HCC were further evaluated using Kaplan-Meier plotter database. Target genes of DE-miRNAs were predicted by miRNet. Then, protein-protein interaction (PPI) networks were established for these targets via the STRING database, after which hub genes in the networks were identified by Cytoscape. Functional annotation and pathway enrichment analyses for the target genes were performed through DAVID database. Three enriched pathways related to HBV-related HCC were selected for further analysis and potential target genes commonly appeared in all three pathways were screened. Cytoscape was employed to construct miRNA-hub gene network. The expression and correlation of potential miRNAs and targets were further detected in clinical HBV-related HCC samples by qRT-PCR.Results7 upregulated and 9 downregulated DE-miRNAs were accessed. 5 of 7 upregulated DE-miRNAs and 5 of 7 downregulated DE-miRNAs indicated significant prognostic roles in HCC. 2312 and 1175 target genes were predicted for the upregulated and downregulated DE-miRNAs, respectively. TP53 was identified as the hub gene in the PPI networks. Pathway enrichment analysis suggested that these predicted targets were linked to hepatitis B, pathways in cancer, microRNAs in cancer and viral carcinogenesis. Further analysis of these pathways screened 20 and 16 target genes for upregulated and downregulated DE-miRNAs, respectively. By detecting the expression of 36 target genes, six candidate target genes were identified. Finally, a potential miRNA-mRNA regulatory network was established based on the results of qRT-PCR and expression correlation analysis.ConclusionsIn the study, potential miRNA-mRNA regulatory pathways were identified, exploring the underlying pathogenesis and effective therapy strategy of HBV-related HCC.