生长激素对感染J亚群禽白血病病毒的鸡胚成纤维细胞系的影响

Effect of Growth Hormone on Chicken Fibroblast Cells Infected with Subgroup J Avian Leukosis Viruses

生长激素(growth hormone,GH)不仅能调节动物的代谢、生长、发育和生殖,也能调节动物的免疫系统。本研究旨在通过转录组测序及生物信息学技术分析GH影响J亚群禽白血病病毒(subgroup J avian leukosis viruses,ALV-J)感染的鸡胚成纤维细胞系(chicken fibroblast cells,DF-1)中的关键基因和信号通路。将空载和GH的过表达质粒分别转染至DF-1,接着均接种ALV-J毒株SCAU-HN06,采用转录组测序技术检测相关的基因表达谱变化。结果显示,与对照组相比,实验组中有上调差异表达基因(differentially expressed genes,DEGs)122个(49%),下调DEGs 127个(51%)。通过GO功能富集分析发现,DEGs主要富集于新陈代谢过程、生物调节、细胞过程、细胞部分、催化活性和结合等过程。经KEGG富集分析发现DEGs主要在免疫系统、信号转导、信号分子和相互作用、细胞生长和死亡等通路上富集。进一步构建蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络,筛选出7个免疫相关基因进行实时定量PCR(RT-qPCR)验证,其中TRAF1、DSTYK、HESX1基因的表达趋势与转录组测序技术(RNA sequencing,RNA-seq)的测序结果相一致。本研究通过生物信息学分析挖掘出可能参与调控ALV-J的关键基因和信号通路,以期为禽白血病病毒-宿主相互作用的关系提供新的见解,为禽白血病的净化提供新的思路。

Growth hormone(GH)not only regulates metabolism,growth,development,and reproduction of animals,but also the immune system of animals.The aim of this study was to analyze key genes and signaling pathways in chicken fibroblast cells(DF-1)infected with subgroup J avian leukosis viruses(ALV-J)after overexpression of GH by transcriptome sequencing and bioinformatics techno-logy.DF-1 was transfected with empty-load plasmid and GH overexpression plasmid respectively,and inoculated with ALV-J strain SCAU-HN06.Transcriptome sequencing was used to detect the changes in the gene expression profile.The results showed that compared with the control group,there were 122(49%)up-regulated differentially expressed genes(DEGs)and 127(51%)down-regulated DEGs in the experimental group.The GO enrichment analysis revealed that DEGs were primarily enriched in me-tabolic processes,biological regulation,cellular processes,cell part,catalytic activity,and binding.The KEGG enrichment analysis found that DEGs were most heavily enriched in the immune system,signal transduction,signaling mole-cular and interaction,cell growth and death pathways.Furthermore,protein-protein interaction(PPI)networks were built,and seven immune-related genes were tested using real-time quantitative PCR(RT-qPCR).The expression trends of TRAF1,DSTYK and HESX1 genes were consistent with the results of RNA sequencing(RNA-seq).In this study,the key genes and signaling pathways that may be involved in the regulation of ALV-J were discovered through bioinformatics analysis.It is expected to provide new insights into the relationship between virus-host interaction in avian leukosis,as well as new ideas for the purification of avian leukosis.