柯萨奇病毒B组5型感染的人恶性胚胎横纹肌肉瘤细胞系lncRNA表达谱分析

LncRNA expression profile in human malignant embryonic rhabdomyosarcoma cell line infected with Coxsackie virus group B type 5

目的探索柯萨奇病毒B组5型(CVB5)感染人恶性胚胎横纹肌肉瘤细胞系(RD)中差异长链非编码RNA(lncRNA)表达谱,为研究CVB5与宿主之间相互作用分子机制提供参考。方法CVB5按感染复数(MOI)为1接种RD细胞24 h后提取总RNA。采用转录组测序技术获取细胞中lncRNA差异表达谱;通过聚类分析、GO分析和KEGG通路富集对差异表达转录本进行了生物信息学分析。同时,利用RNAfold软件对lncRNA进行了二级结构预测。结果与对照组相比,CVB5感染RD细胞后,共有1754个mRNAs和508个lncRNA呈上调表达,3106个mRNAs和760个lncRNA呈下调表达。差异表达lncRNA的共表达基因主要富集在分子结构活性、蛋白质分子结合和体液免疫反应等生物过程;lncRNA靶基因主要参与嗅觉传导途径、细胞因子受体相互作用和神经活性配体受体相互作用等通路。此外,实时荧光定量PCR验证的7个差异表达lncRNA与测序结果一致。结论CVB5感染RD细胞后,差异显著性lncRNA主要参与了免疫相关过程,为充分理解lncRNA在CVB5感染中的调控作用奠定了基础。

Objective To explore the molecular mechanism of the interaction between Coxsackie virus group B type 5(CVB5)and the host,The long non-coding RNA(lncRNA)expression profile in human malignant embryonic rhabdomyosarcoma cell line(RD)infected by CVB5 was investigated.Methods After 24 hours of infection with 1MOI CVB5 RNA was sampled and RNA-sequencing was used to identify the differential expression profile of lncRNA in the cells.Differentially expressed transcripts were analyzed by cluster analysis,GO analysis and KEGG pathway enrichment.At the same time,RNAfold was used to predict the secondary structure of lncRNA.Results Compared to the uninfected group,1754 mRNAs and 508 lncRNAs were up-regulated and 3106 mRNAs/760 lncRNAs were down-regulated after CVB5 infection.Co-expressed genes that differentially express lncRNA were mainly enriched in biological processes such as molecular structure activity,protein molecular binding,and humoral immune response;lncRNA targets were mainly involved in olfactory transduction pathway,cytokine-cytokine receptors interaction,neuroactive ligand-receptor interaction and other pathways.In addition,the seven differen-tially expressed lncRNAs verified by quantitative real-time PCR(RT-qPCR)were consistent with the sequencing results.Conclusions lncRNA is mainly involved in the regulation of the immune processes,which lays the foundation for a full understanding of the regulatory role of lncRNA in CVB5 infection.