摘要
目的 探讨人脐带血间充质干细胞(hUCB-MSCs)治疗肝纤维化microRNA(miRNA)表达的变化及其功能.方法 从GEO数据库获取肝星形细胞(LX-2)与hUCB-MSCs共培养后测序的miRNA表达矩阵,筛选出差异表达miRNA(DEMs);利用miRWalk在线数据库预测DEMs的靶基因,并通过Cytoscape对miRNA-mRNA网络进行可视化;通过GeneMANIA数据库构建DEMs靶基因的蛋白质互作网络,并对DEMs的靶向基因进行功能和通路富集分析.结果 差异表达分析显示,与hUCB-MSCs共培养的LX-2细胞较单独培养的LX-2细胞存在13个低表达DEMs和28个高表达DEMs.通过构建miRNA-mRNA网络分析显示,hsa-miR-221-3p及hsa-miR-146a-5p同时靶向ERBB4,hsa-miR-10a-5p及hsa-miR-155-5p同时靶向RORA.GO分析发现,这些DEMs靶向基因主要富集的生物过程包括GDP折叠、蛋白磷酸酶调节活性、核受体活性、配体激活的转录因子活性及纤维蛋白原连接等.KEGG分析发现,这些DEMs靶向基因主要在癌症miRNA、癌症蛋白聚糖类、肝癌、Rap1信号通路、胃癌、甲状腺癌以及膀胱癌方面富集.结论 本研究确定了 41种DEMs及ERBB4、RORA两种多miRNA共同靶向基因,它们可能在hUCB-MSC治疗肝纤维化的过程中发挥作用.
Abstract
Objective To investigate the changes in the expression of microRNAs(miRNAs)and their functions after the treatment of liver fibrosis using human umbilical cord blood mesenchymal stem cells(hUCB-MSCs).Methods The GEO data-base was used to obtain the expression matrix of miRNAs sequenced after co-culture of human hepatic stellate cells LX-2 and hUCB-MSCs,and differentially expressed miRNAs(DEMs)were identified.The miRWalk online database was used to predict the target genes of DEMs,and Cytoscape was used to visualize the miRNA-mRNA network.The GeneMANIA database was used to construct a protein-protein interaction network for the target genes of DEMs,and then functional and pathway enrichment analyses were performed for the target genes of DEMs.Results The differential analysis showed that compared with the LX-2 cells cul-tured alone,the LX-2 cells co-cultured with hUCB-MSCs had 13 downregulated DEMs and 28 upregulated DEMs.The miRNA-mRNA network analysis showed that both hsa-miR-221-3p and hsa-miR-146a-5p targeted ERBB4,and both hsa-miR-10a-5p and hsa-miR-155-5p targeted RORA.The GO analysis showed that the target genes of the DEMs were mainly enriched in the biological processes such as GDP folding,protein phosphatase regulator activity,nuclear receptor activity,ligand-activated transcription fac-tor activity,and fibrinogen binding,and the KEGG analysis showed that these genes were mainly enriched in the pathways asso-ciated with cancer miRNAs,cancer proteoglycans,liver cancer,the Rap1 signaling pathway,gastric cancer,thyroid cancer,and bladder cancer.Conclusion This study identifies 41 DEMs and 2 target genes,ERBB4 and RORA,of multiple miRNAs,which may play a role in the treatment process of liver fibrosis by hUCB-MSCs.
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