Role of WDHD1 in Human Papillomavirus-Mediated Oncogenesis Identified by Transcriptional Profiling of E7-Expressing Cells.
Role of WDHD1 in Human Papillomavirus-Mediated Oncogenesis Identified by Transcriptional Profiling of E7-Expressing Cells.
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通过 E7 表达细胞的转录谱鉴定 WDHD1 在人乳头瘤病毒介导的肿瘤发生中的作用。
DOI：
10.1128/jvi.00513-16
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发表时间：
2016
影响因子：
5.4
通讯作者：
Chen Jason J
中科院分区：
文献类型：
--
作者：
Zhou Yunying;Zhang Qishu;Gao Ge;Zhang Xiaoli;Liu Yafei;Yuan Shoudao;Wang Xiaowei;Chen Jason J
高危型人乳头瘤病毒（HPV）的E7癌蛋白在HPV诱导的癌发生中起主要作用。E7废除G1细胞周期检查点并诱导基因组不稳定性，但其机制尚未完全了解。在这项研究中，我们进行了RNA测序（RNA-seq），以表征表达HPV 16（HPV-16）E7的角质形成细胞的转录谱。在转录组水平上，236个基因在E7和载体对照细胞之间差异表达。通过实时PCR进一步证实了差异表达基因的一个子集，其中大多数是E7表达细胞的新基因。有趣的是，多种转录因子的活性在E7表达细胞中改变。通过生物信息学分析，研究了E7表达细胞中改变的途径。上调的基因在细胞周期和DNA复制中富集，以及在DNA代谢过程、转录、DNA损伤、DNA修复和核苷酸代谢中富集。具体来说，我们将我们的研究集中在编码WDHD 1（WD重复和高迁移率族[HMG]盒DNA结合蛋白）的基因上，这是E7表达细胞中上调的基因之一。WDHD 1是调节DNA复制的复制体的组分。最近的研究表明，WDHD 1也可能作为一个DNA复制起始因子以及G1检查点调节器。我们发现，在E7表达细胞中，WDHD 1蛋白的稳态水平随着半衰期的增加而沿着增加。此外，下调WDHD 1减少E7诱导的G1检查点废除和再复制，证明了WDHD 1的新功能。这些研究揭示了HPV诱导基因组不稳定性的机制，并具有治疗意义。重要信息高危型HPV诱导宫颈癌，并编码E7癌蛋白，在HPV诱导的致癌作用中起着重要作用。然而，E7诱导致癌的机制尚未完全了解;没有特异性抗HPV药物。在这项研究中，我们进行了RNA-seq来表征表达HPV-16 E7的角质形成细胞的转录谱，并确定了E7和载体对照细胞之间差异表达的200多个基因。通过生物信息学分析，鉴定了E7表达细胞中改变的途径。值得注意的是，WDHD 1基因，在E7表达细胞中上调的基因之一，被发现在E7诱导的G1检查点废除和再复制中发挥重要作用。这些研究揭示了HPV诱导基因组不稳定性的机制，并具有治疗意义。
The E7 oncoprotein of the high-risk human papillomavirus (HPV) plays a major role in HPV-induced carcinogenesis. E7 abrogates the G1cell cycle checkpoint and induces genomic instability, but the mechanism is not fully understood. In this study, we performed RNA sequencing (RNA-seq) to characterize the transcriptional profile of keratinocytes expressing HPV 16 (HPV-16) E7. At the transcriptome level, 236 genes were differentially expressed between E7 and vector control cells. A subset of the differentially expressed genes, most of them novel to E7-expressing cells, was further confirmed by real-time PCR. Of interest, the activities of multiple transcription factors were altered in E7-expressing cells. Through bioinformatics analysis, pathways altered in E7-expressing cells were investigated. The upregulated genes were enriched in cell cycle and DNA replication, as well as in the DNA metabolic process, transcription, DNA damage, DNA repair, and nucleotide metabolism. Specifically, we focused our studies on the gene encoding WDHD1 (WD repeat and high mobility group [HMG]-box DNA-binding protein), one of the genes that was upregulated in E7-expressing cells. WDHD1 is a component of the replisome that regulates DNA replication. Recent studies suggest that WDHD1 may also function as a DNA replication initiation factor as well as a G1checkpoint regulator. We found that in E7-expressing cells, the steady-state level of WDHD1 protein was increased along with the half-life. Moreover, downregulation of WDHD1 reduced E7-induced G1checkpoint abrogation and rereplication, demonstrating a novel function for WDHD1. These studies shed light on mechanisms by which HPV induces genomic instability and have therapeutic implications.IMPORTANCEThe high-risk HPV types induce cervical cancer and encode an E7 oncoprotein that plays a major role in HPV-induced carcinogenesis. However, the mechanism by which E7 induces carcinogenesis is not fully understood; specific anti-HPV agents are not available. In this study, we performed RNA-seq to characterize transcriptional profiling of keratinocytes expressing HPV-16 E7 and identified more than 200 genes that were differentially expressed between E7 and vector control cells. Through bioinformatics analysis, pathways altered in E7-expressing cells were identified. Significantly, the WDHD1 gene, one of the genes that is upregulated in E7-expressing cells, was found to play an important role in E7-induced G1checkpoint abrogation and rereplication. These studies shed light on mechanisms by which HPV induces genomic instability and have therapeutic implications.