Department of Medical Microbiology, Faculty of Health Sciences, University of Manitoba, Winnipeg, MB R3E 0W2, Canada.
Cadham Provincial Laboratory, Manitoba Health, Winnipeg, MB R3C 3J7, Canada.
Viruses. 2022 Aug 12;14(8):1764. doi: 10.3390/v14081764.
The high-risk human papillomaviruses are oncogenic viruses associated with almost all cases of cervical carcinomas, and increasing numbers of anal, and oral cancers. Two oncogenic HPV proteins, E6 and E7, are capable of immortalizing keratinocytes and are required for HPV associated cell transformation. Currently, the influence of these oncoproteins on the global regulation of the host proteome is not well defined. Liquid chromatography coupled with quantitative tandem mass spectrometry using isobaric-tagged peptides was used to investigate the effects of the HPV16 oncoproteins E6 and E7 on protein levels in human neonatal keratinocytes (HEKn). Pathway and gene ontology enrichment analyses revealed that the cells expressing the HPV oncoproteins have elevated levels of proteins related to interferon response, inflammation and DNA damage response, while the proteins related to cell organization and epithelial development are downregulated. This study identifies dysregulated pathways and potential biomarkers associated with HPV oncoproteins in primary keratinocytes which may have therapeutic implications. Most notably, DNA damage response pathways, DNA replication, and interferon signaling pathways were affected in cells transduced with HPV16 E6 and E7 lentiviruses. Moreover, proteins associated with cell organization and differentiation were significantly downregulated in keratinocytes expressing HPV16 E6 + E7. High-risk HPV E6 and E7 oncoproteins are necessary for the HPV-associated transformation of keratinocytes. However their influence on the global dysregulation of keratinocyte proteome is not well documented. Here shotgun proteomics using TMT-labeling detected over 2500 significantly dysregulated proteins associated with E6 and E7 expression. Networks of proteins related to interferon response, inflammation and DNA damage repair pathways were altered.
高危型人乳头瘤病毒是致癌病毒，与几乎所有宫颈癌病例以及越来越多的肛门癌和口腔癌有关。两种致癌 HPV 蛋白 E6 和 E7 能够使角质形成细胞永生化，并且是 HPV 相关细胞转化所必需的。目前，这些致癌蛋白对宿主蛋白质组的全球调控的影响还没有很好地定义。使用同位素标记肽的液相色谱-串联质谱定量技术，研究了 HPV16 致癌蛋白 E6 和 E7 对人新生儿角质形成细胞（HEKn）中蛋白质水平的影响。途径和基因本体富集分析显示，表达 HPV 致癌蛋白的细胞中与干扰素反应、炎症和 DNA 损伤反应相关的蛋白质水平升高，而与细胞组织和上皮发育相关的蛋白质水平下调。这项研究鉴定了与 HPV 致癌蛋白在原代角质形成细胞中失调的途径和潜在的生物标志物，这可能具有治疗意义。值得注意的是，转导 HPV16 E6 和 E7 慢病毒的细胞中受影响的途径包括 DNA 损伤反应途径、DNA 复制和干扰素信号通路。此外，表达 HPV16 E6 + E7 的角质形成细胞中与细胞组织和分化相关的蛋白质显著下调。高危型 HPV E6 和 E7 致癌蛋白是 HPV 相关角质形成细胞转化所必需的。然而，它们对角质形成细胞蛋白质组全局失调的影响尚未得到很好的记录。在这里，使用 TMT 标记的鸟枪法蛋白质组学检测到与 E6 和 E7 表达相关的超过 2500 个显著失调的蛋白质。与干扰素反应、炎症和 DNA 损伤修复途径相关的蛋白质网络发生了改变。
