人正常宫颈上皮细胞感染沙眼衣原体的基因表达谱分析

Gene expression profile in human cervical epithelial cells infected with Chlamydia trachomatis

目的分析人正常宫颈上皮细胞(HcerEpic)感染沙眼衣原体(Chlamydia trachomatis,Ct)前后基因表达谱差异。方法HcerEpic细胞经DEAE-D预处理后加入E型Ct标准株,培养44 h后收集HcerEpic细胞(感染组),并以未感染Ct的HcerEpic细胞作为对照组,提取两组总RNA后反转录并构建cDNA文库。通过高通量测序分析两组基因表达谱差异,并选择代表性基因通过实时荧光定量PCR(qPCR)验证。结果共检测23997个基因,差异基因125个,其中上调基因119个,下调基因6个;GO分析显示差异基因富集于对病毒防御反应、Ⅰ型干扰素(interferon,IFN)信号通路、对Ⅰ型IFN的细胞反应等条目;KEGG富集通路为甲型流感、单纯疱疹病毒感染、EB病毒感染、HPV感染、NOD样受体通路等相关通路。结论人正常宫颈上皮细胞感染Ct前后转录组存在差异,差异基因主要富集于IFN通路,与细胞抗病毒过程密切相关。qPCR验证了ISG15、IFIT2、OASL、UBE2L6等差异显著且与IFN通路密切相关的基因。

Objective To compare gene expression profiles in normal human cervical epithelial cells(HcerEpic)before and after Chlamydia trachomatis(Ct)infection.Methods HcerEpic cells that were pretreated with DEAE-D were infected with Ct serotype E standard strain and then cultured for 44 h.Uninfected HcerEpic cells were used as the control group.Total RNA was extracted from the cells in each group and reverse transcribed to construct a cDNA library.Differences in gene expression profiles between the two groups were analyzed by high-throughput sequencing and the representative genes were selected for verification by qPCR.Results A total of 23997 genes were detected,including 125 differentially expressed genes.Among the 125 genes,119 were up-regulated and six were down-regulated.GO analysis showed that the differentially expressed genes were enriched in several biological processes including defense response to virus,typeⅠinterferon signaling pathway and cellular responses to typeⅠinterferons.KEGG enrichment analysis showed the differentially expressed genes were mainly enriched in the pathways related to virus infections,such as influenza A virus,herpes simplex virus,EB virus and HPV,and NOD-like receptor pathway.Conclusions There were significant differences in transcriptome profiles of HcerEpic cells before and after Ct infection.The differentially expressed genes were mainly enriched in the interferon pathway,which was closely related to the antiviral processes in cells.qPCR verified the differentially expressed genes and the genes closely related to the interferon pathway,such as ISG15,IFIT2,OASL and UBE2L6.