BHV-1感染MDBK细胞lncRNA表达谱系及lncRNA与mRNA的关联性分析  

The expression profile of LncRNA and correlation analysis between lncRNAs and mRNA in BHV-1 infected MDBK cells

作　　者：黄艳梅 白永飞 黄麒霏 马金柱[1] 宋佰芬[1] HUANG Yan-mei;BAI Yong-fei;HUANG Qi-fei;MA Jin-zhu;SONG Bai-fen(College of Life Science and Technology,Heilongjiang Bayi Agricultural University,Daqing 163319,China)

机构地区：[1]黑龙江八一农垦大学生命科学技术学院,黑龙江大庆163319

出　　处：《中国预防兽医学报》2021年第9期930-938,共9页Chinese Journal of Preventive Veterinary Medicine

基　　金：“十三五”国家重点研发计划(牛羊常见病毒病的诊断与检测新技术研究/2016YFD0500904)。

摘　　要：为探究牛疱疹病毒1型(BHV-1)感染MDBK细胞的长链非编码RNA(lncRNA)表达谱系变化及lncRNA与m RNA的关联性,本研究将BHV-1感染MDBK细胞,以未感染BHV-1的MDBK细胞作为对照,于感染后采用Illumina HiSeq^(TM)4000测序,测序数据基于stringtie进行转录本重构,得到7 935个lncRNA转录本,再用CPC和CNCI两个软件进行新转录本编码能力的预测,获得681个新产生的lncRNA转录本。利用DESeq2软件对差异表达的lncRNA进行分析,结果显示,实验组共获得839个差异表达的lncRNA转录本,其中表达上调转录本508个,表达下调转录本331个。利用RNA plex软件对反义lncRNA与mRNA进行关联性分析,得到lncRNAmRNA相互作用的靶基因对1 227个,其中差异表达的lncRNA-mRNA相互作用靶基因对42个;顺式作用lncRNA与m RNA关联分析,获得lncRNA-mRNA靶基因对3 793个,差异表达的lncRNA-mRNA靶基因对149个;反式作用lncRNA与mRNA关联分析,获得lncRNA-mRNA靶基因对268 409个。通过基因本体论(GO)注释和京都基因与基因组百科全书(KEGG)信号通路富集分析进一步对与lncRNA关联mRNA的功能和途径进行分析,结果显示,关联的mRNA在细胞进程、单组织代谢过程、代谢过程及生物学调节等过程显著富集。随机选取10个差异表达的lncRNA,利用qRT-PCR方法进行验证,结果显示,其表达变化趋势与高通量测序数据趋势一致。本研究为进一步研究lncRNA在病毒感染中的作用奠定了基础,同时为进一步阐述BHV-1的发病机制、致病机理提供参考。In order to investigate the changes of lncRNA expression profile and its correlation with mRNA in BHV-1 infected MDBK cells,MDBK cells were infected with BHV-1,samples were collected at different times and sequenced with Illumina HiSeq^(TM) 4000.Sequencing data were reconstructed based on stringtie to obtain the total lncRNA.CPC and CNCI software were used to predict the coding ability of new transcripts.7935 lncRNA transcripts were obtained,among which 7255 were known and 681 were newly generated.DESeq2 software was used to analyze the differentially expressed lncRNAs,and the results showed that 839 differentially expressed lncRNA transcripts were obtained in the experimental group,including 508 up-regulated and 331 down-regulated transcripts.The association between antisense lncRNAs and mRNAs was analyzed by RNA plex software,and 1227 target gene pairs of lncRNA-mRNA interaction were obtained,among which 42 were differentially expressed.Correlation analysis of cis lncRNAs and mRNAs showed that 3793 pairs of lncRNA-mRNA target genes were obtained and 149 pairs of lncRNA-mRNA target genes were differentially expressed.Correlation analysis of trans lncRNA and mRNA revealed 268409 target gene pairs of lncRNA-mRNA.The functions and pathways of lncRNA-associated mRNAs were further analyzed by GO annotation and Kyoto KEGG signaling pathway enrichment analysis.The results showed that mRNAs were significantly enriched in cellular processes,single-tissue metabolic processes,metabolic processes,and biological regulation.Ten differentially expressed lncRNAs were randomly selected and verified by qRT-PCR.The results showed that the expression trend was consistent with the high-throughput sequencing data.This study lays a foundation for further study of the role of lncRNA in viral infection.It provides a reference for elucidation of the pathogenesis of BHV-1.

关 键 词：牛疱疹病毒1型 MDBK细胞 差异表达 LncRNA 转录组测序 关联性分析 

分 类 号：S855.3[农业科学—临床兽医学]