鉴定潜在的miRNA-mRNA调节网络有助于HBV相关HCC的发病机制

StephenW

Identification of potential miRNA–mRNA regulatory network contributing to pathogenesis of HBV-related HCC

    Weiyang Lou†, Jingxing Liu†, Bisha Ding, Danni Chen, Liang Xu, Jun Ding, Donghai Jiang, Lin Zhou, Shusen ZhengEmail author and Weimin FanEmail author

†Contributed equally
Journal of Translational Medicine201917:7

https://doi.org/10.1186/s12967-018-1761-7

©  The Author(s) 2019

    Received: 22 October 2018Accepted: 21 December 2018Published: 3 January 2019

Abstract
Background

Hepatitis 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.
Methods

Microarray 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.
Results

7 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.
Conclusions

In the study, potential miRNA–mRNA regulatory pathways were identified, exploring the underlying pathogenesis and effective therapy strategy of HBV-related HCC.

StephenW

鉴定潜在的miRNA-mRNA调节网络有助于HBV相关HCC的发病机制

    Weiyang Lou†，Jingxing Liu†，Bisha Ding，Danny Chen，Liang Xu，Jun Ding，Donghai Jiang，Lin Zhou，Shusen ZhengEmail author and Weimin FanEmail author

†同等贡献
转化医学杂志201917：7

https://doi.org/10.1186/s12967-018-1761-7

©作者2019

    收稿日期：2018年10月22日接受日期：2018年12月21日出版日期：2019年1月3日

抽象
背景

乙型肝炎病毒（HBV）是肝细胞癌（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HCC。然而，仍然缺乏HBV相关HCC中的综合miRNA-mRNA调节网络。本研究旨在鉴定有助于HBV相关HCC发病机制的潜在miRNA-mRNA调节途径。
方法

微阵列GSE69580从Gene Expression Omnibus（GEO）数据库下载。使用GEO2R和'R-limma'进行差异表达分析。在两个分析组中出现的常见miRNA被筛选为潜在的差异表达的miRNA（DE-miRNA）。使用Kaplan-Meier绘图仪数据库进一步评估筛选的DE-miRNA在HCC中的预后作用。 miRNet预测了DE-miRNA的靶基因。然后，通过STRING数据库为这些靶标建立蛋白质 - 蛋白质相互作用（PPI）网络，之后通过Cytoscape鉴定网络中的中枢基因。通过DAVID数据库进行靶基因的功能注释和途径富集分析。选择与HBV相关的HCC相关的三种富集途径用于进一步分析，并筛选通常出现在所有三种途径中的潜在靶基因。 Cytoscape用于构建miRNA-hub基因网络。通过qRT-PCR在临床HBV相关HCC样品中进一步检测潜在miRNA和靶标的表达和相关性。
结果

获得了7种上调的和9种下调的DE-miRNA。 7种上调的DE-miRNA中的5种和7种下调的DE-miRNA中的5种表明在HCC中具有显着的预后作用。分别预测了上调和下调的DE-miRNA的2312和1175个靶基因。 TP53被确定为PPI网络中的中枢基因。通路富集分析表明，这些预测的靶标与乙型肝炎，癌症途径，癌症中的微小RNA和病毒致癌作用有关。对这些途径的进一步分析分别针对上调和下调的DE-miRNA筛选20和16个靶基因。通过检测36个靶基因的表达，鉴定了6个候选靶基因。最后，基于qRT-PCR和表达相关性分析的结果建立了潜在的miRNA-mRNA调节网络。
结论

在该研究中，确定了潜在的miRNA-mRNA调节途径，探索了HBV相关HCC的潜在发病机制和有效治疗策略。

StephenW

https://translational-medicine.b ... 6/s12967-018-1761-7