miR-130a-3p在胶质母细胞瘤中的表达与靶基因分析

Expression and Target Genes Analysis of miR-130a-3p in Glioblastoma

目的探索miR-130a-3p在胶质母细胞瘤(glioblastoma,GBM)细胞中的表达水平,预测并分析miR-130a-3p的靶基因。方法采用实时聚合酶链反应(polymerase chain reaction,PCR)检测GBM细胞和正常人胶质细胞中miR-130a-3p的相对表达水平。选择miRanda、DIANA、TargetScan和PicTar工具预测miR-130a-3p的靶基因,对靶基因进行基因本体(gene ontology,GO)富集分析和通路富集分析,基于STRING数据库进一步建立蛋白互作网络(protein-protein interaction,PPI)并利用Cytoscape软件进行可视化分析。结果相对于正常人胶质细胞,miR-130a-3p在GBM细胞中的表达水平显著下调。四种预测工具取交集后共筛选出302个靶基因。GO富集分析表明miR-130a-3p靶基因主要参与转录调控、蛋白泛素化、Wnt信号、miRNA介导基因沉默、DNA损伤反应、细胞迁移等生物学过程和分子功能。通路富集分析发现miR-130a-3p靶基因主要富集于转化生长因子β(transforming growth factor-β,TGF-β)信号通路、磷脂酰肌醇信号通路、内吞、自噬调节、胶质瘤、FoxO信号通路等。结论miR-130a-3p在GBM细胞中低表达,其靶基因涉及多个肿瘤发生相关的信号通路,为进一步阐明其在GBM发生过程中的调控机制以及开发靶向治疗提供参考。

Objective To investigate the expression level of miR-130 a-3 p in glioblastoma(GBM)and predict the target genes of miR-130 a-3 p.Methods The relative expression levels of miR-130 a-3 p in GBM cell lines and normal human astrocyte cell line were detected by real-time polymerase chain reaction(PCR).The target genes of miR-130 a-3 p were predicted using miRanda,DIANA,TargetScan and PicTar tools,then gene ontology(GO)enrichment and pathway enrichment analysis were performed to study the function of predicted target genes.The protein-protein interaction(PPI)network for target genes was constructed by STRING database and the visualization was analysed by Cytoscape software.Results Compared with the normal human astrocyte cell line,the expression level of miR-130 a-3 p in GBM cell lines were significantly downregulated.A total of 302 target genes were obtained from the intersection of four prediction tools of miRNA targets.GO enrichment analysis showed that the target gene of miR-130 a-3 p were mainly participated in the biological processes and molecular functions such as regulation of transcription,protein polyubiquitination,Wnt signaling pathway,gene silencing by miRNA,DNA damage response,and cell migration.Pathway enrichment analysis found that target gene of miR-130 a-3 p were significantly enriched in transtorming growth factor-β(TGF-β)signaling pathway,phosphatidylinositol signaling system,endocytosis,regulation of autophagy,glioma and FoxO signaling pathway.Conclusion The expression of miR-130 a-3 p is downregulated in GBM cell and its target genes are involved in multiple biological processes and signaling pathways associated with tumorigenesis,which provides references for further clarify its regulatory mechanism and development of targeted therapy for GBM.