羊口疮病毒感染对山羊皮肤成纤维细胞circRNA表达谱的影响

The influence of orf virus infection on circular RNA expression profile in goat skin fibroblast cells

旨在探究羊口疮病毒(orf virus,ORFV)感染对山羊皮肤成纤维细胞(goat skin fibroblast,GSF)环状RNA(circular RNA,circRNA)表达谱的影响。以ORFV感染的GSF和未感染细胞为对象构建6个cDNA文库进行转录组测序,利用生物信息学方法进行circRNA鉴定、circRNA差异表达分析、差异circRNA的GO和KEGG功能富集分析,最后挑选9个差异circRNA进行qPCR和Sanger测序验证。结果发现,在未感染组和感染组分别鉴定到9979和10844个circRNA,共有151个circRNA差异表达,其中59个circRNA表达上调,92个circRNA表达下调;GO富集分析表明,差异circRNA主要富集在炎症应答、上皮结构维持、细胞迁移正调控、泛素蛋白转移酶活性等生物学过程;KEGG富集分析表明,差异circRNA主要富集在紧密连接、黏着斑、血管平滑肌收缩、内吞作用、细胞因子受体相互作用等信号通路;circRNA1001,circRNA1684,circRNA3127和circRNA788表达量与转录组测序结果一致。本研究首次报道了ORFV感染对GSF细胞circRNA表达谱的影响及差异circRNA在ORFV感染中的潜在功能,为ORFV与宿主互作机制研究提供新的线索。

The aim of this study was to explore the influence of orf virus(ORFV)infection on circular RNA(circRNA)expression profile in goat skin fibroblast(GSF)cells.ORFV-infected GSF cells and uninfected samples were used to construct six cDNA libraries for transcriptome sequencing.Bioinformatics methods were used to perform circRNA identification,differential expression analysis of circRNAs and Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.Furthermore,nine circRNAs were selected for qPCR and Sanger sequencing validation.The results indicated that a total of 9979and 10844circRNAs were identified in GSF group and OV group,respectively.151circRNAs were differentially expressed(DE)with59upregulated and 92downregulated following ORFV infection.GO enrichment analysis indicated that DE circRNAs were significantly enriched in biological processes such as regulation of inflammatory response,epithelial structure maintenance,positive regulation of cell migration,positive regulation of ubiquitin-protein transferase activity,etc.KEGG enrichment analysis indicated that DE circRNAs were significantly enriched in tight junction,focal adhesion,vascular smooth muscle contraction,endocytosis and cytokine-cytokine receptor interaction,etc.Furthermore,circRNA1001,circRNA1684,circRNA3127and circRNA7880were successfully validated,consistent with the high-throughput sequencing results.The study first reported the influence of ORFV infection on circRNA expression profile in GSF cells and the potential function of DE circRNAs during ORFV infection,providing new clues for the mechanism of ORFV and host interaction.