人疱疹病毒6型感染HSB-2细胞的lncRNA表达谱变化分析

Changes in expression profile of lncRNA in HSB-2 cells infected by human herpesvirus 6

目的运用高通量测序技术检测人疱疹病毒6型(human herpesvirus 6,HHV-6)感染人淋巴细胞系HSB-2后长链非编码RNA(long non-coding RNA,lncRNA)表达谱的改变,探究lncRNA在HHV-6感染复制中的作用。方法HHV-6感染HSB-2细胞72 h后,提取对照细胞和病毒感染细胞的RNA进行测序,筛选差异表达的lncRNA;通过生物信息学方法对预测的lncRNA靶基因进行GO注释和KEGG信号通路分析,构建共表达网络图。qRT-PCR检测差异表达lncRNA的变化倍数。结果共筛选到612个显著差异表达lncRNA,其中420个为表达上调,192个表达下调。通过对lncRNA靶基因进行GO和KEGG富集分析显示,差异表达lncRNA的靶基因与表观染色体调控、免疫应答及细胞代谢等生物学过程密切相关。qRT-PCR确定10条上调IncRNAs表达变化趋势与高通量测序数据一致。结论本研究对HHV-6感染HSB-2细胞的lncRNA表达谱进行分析,为深入探索lncRNA在HHV-6复制增殖及相关疾病中的作用奠定基础。

Objective To examine the changes in long noncoding RNA(lncRNA)expression in human lymphocyte cells infected with human herpesvirus 6(HHV-6)using high throughput sequencing,so as to explore the effects of lncRNA in HHV-6 infection and replication.Methods After 72 hours infection of HSB-2 cells by HHV-6,RNA from control cell and virus infected cells was sequenced to screen differentially expressed lncRNA.GO annotation and signal pathway analysis for the predicted target genes of lncRNA were performed and co-expression network map was constructed by bioinformatic methods.qRT-PCR method was used to detect the multiple times of differential expression of lncRNA.Results A total of 612 significantly differentially expressed lncRNA were obtained,including 420 of up regulation and 192 of down regulation.GO analysis and KEGG enrichment analysis for the target genes of lncRNA indicated that target genes of the differentially expressed IncRNAs involved in the biological processes of epigenic regulation of chromosome,immune response and cell metabolism.The trends of expression of 10 differentially expressed lncRNA of up regulation verified by quantitative real time PCR were in consistent with high throughput sequencing data.Conclusions The current study analyzed differentially expression of lncRNA in HSB-2 cells infected with HHV-6,and provided basis for further exploration of the role of lncRNA in HHV-6 pathogenesis.