Human gene expression microarray analysis of the HPV 6bE7-HaCaT stable cell line.
Human gene expression microarray analysis of the HPV 6bE7-HaCaT stable cell line.
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DOI：
10.1016/j.gene.2018.02.067
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发表时间：
2018-05
期刊：
影响因子：
3.5
通讯作者：
Boya Zhang;Xianzhen Chen;Qiang Zhou;Yin-jing Song;Siyuan Sun;Hao Cheng
中科院分区：
文献类型：
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作者：
Boya Zhang;Xianzhen Chen;Qiang Zhou;Yin-jing Song;Siyuan Sun;Hao Cheng
目的人乳头瘤病毒(HPV)是世界上最常见的性传播媒介。HPV6b是一种低风险类型的HPV，会导致皮肤和肛门生殖器粘膜的良性疣状增生性病变。以前的研究表明，HPV基因6型有时与高级别病变和肛门癌有关。目前，低危型HPV感染的发病机制及其与高危型HPV的关系尚不清楚。HPV早期表达基因编码的E7蛋白在HPV感染中起重要作用。本研究的目的是研究HPV6b E7转染人HaCaT稳定细胞系的人基因表达特征。对HPV6b感染差异表达基因的鉴定可能有助于我们更全面地了解HPV6b感染，并将有助于我们探索E7蛋白的特定作用。方法建立稳定的HPV6b E7基因转染细胞系，并用基因芯片分析该细胞系的全基因组表达谱。采用实时定量聚合酶链式反应(qRT-PCR)对差异表达基因进行验证。采用GO富集法对基因进行功能注释。结果HPV6bE7-HaCaT稳定细胞系与对照细胞系共鉴定出3519个差异表达基因，其中1 884个基因表达上调，1635个基因表达下调，两组间 >； 2.0发生折叠变化。定量逆转录聚合酶链式反应分析HPV6bE7-HaCaT稳定细胞系中前20个上调基因和前20个下调基因的表达谱与芯片数据一致。HPV6bE7-HaCaT细胞中HPV6bE7-HaCaT基因表达增强最明显的是SIMC1、S100A8和S100A9，而PXDN的表达显著下调。GO分析显示，HPV6bE7主要影响生物学过程，最显著的差异是心脏诱导(GO：0003129)。许多差异表达基因与组蛋白H4-K20去甲基化(GO：0035574)相关。KEGG分析显示，HPV6bE7-HaCaT稳定细胞系的基因表达变化最显著的是初级胆汁酸的生物合成，最多样的生物学过程与系统性红斑狼疮的发病有关。结论分析了HPV6bE7-HaCaT稳定细胞系的整体基因表达谱，揭示了E7蛋白调控的基因，为HPV6b感染的发病机制提供了新的思路。
ObjectiveHuman papillomavirus (HPV) is the most common sexually transmitted agent in the world. HPV6b is a low-risk type of HPVs that causes benign verrucous hyperplastic lesions of the skin and anal genital mucosa. Previous research has indicated that HPV genotype 6 is sometimes associated with high-grade lesions and anal cancer. The pathogenesis of low-risk HPV infection and its relationship to high-risk HPV is not clear at present. The E7 protein, which is encoded by HPV early –expressing genes, plays an important role in HPV infection. The aim of this study is to investigate the human gene expression signature of the HPV6b E7-transfected HaCaT stable cell line. The identification of differentially expressed genes might provide a more comprehensive understanding of HPV6b infection and will allow us to explore the specific role of E7 protein.MethodsWe established a stable cell line transfected with the HPV6b E7 gene and analyzed the line's genome-wide expression profile by microarray. Quantitative real-time PCR (qRT-PCR) was used to verify the differentially expressed genes. GO enrichment analysis was applied for gene annotation according to functions. KEGG analysis, a system for analyzing gene function and genome information, was used to help us integrate differentially expressed genes into pathways.ResultsA total of 3519 genes were identified to be significantly differentially expressed between the HPV 6bE7-HaCaT stable cell line and a control cell line, among which 1884 genes were up-regulated and 1635 genes were down-regulated with a fold-change > 2.0 between the two groups. The expression profiles of the top 20 up-regulated and the top 20 down-regulated genes in the HPV 6bE7-HaCaT stable cell line as analyzed by qRT-PCR were consistent with the microarray data. The most significantly enhanced genes HPV 6bE7-HaCaT cells were SIMC1, S100A8 and S100A9, whereas PXDN expression was markedly down-regulated. GO analysis showed that HPV 6bE7 primarily affected biological processes and that the most significant difference was in heart induction (GO:0003129). Many differentially expressed genes were linked to histone H4-K20 demethylation (GO:0035574). KEGG analysis showed that the most significant changes in gene expression were related to primary bile acid biosynthesis, and the most diverse biological processes were related to systemic lupus erythematosus pathogenesis.ConclusionsThe global gene expression profile of the HPV 6bE7-HaCaT stable cell line was analyzed, revealing the genes regulated by E7 protein and providing insight into the pathogenesis of HPV6b infection.