Harden Mallory E, Prasad Nripesh, Griffiths Anthony, Munger Karl
Program in Virology, Division of Medical Sciences, Harvard Medical School, Boston, Massachusetts, USA.
Department of Developmental, Molecular and Chemical Biology, Tufts University School of Medicine, Boston, Massachusetts, USA.
mBio. 2017 Jan 3;8(1):e02170-16. doi: 10.1128/mBio.02170-16.
The E6 and E7 proteins are the major oncogenic drivers encoded by high-risk human papillomaviruses (HPVs). While many aspects of the transforming activities of these proteins have been extensively studied, there are fewer studies that have investigated how HPV E6/E7 expression affects the expression of cellular noncoding RNAs. The goal of our study was to investigate HPV16 E6/E7 modulation of cellular microRNA (miR) levels and to determine the potential consequences for cellular gene expression. We performed deep sequencing of small and large cellular RNAs in primary undifferentiated cultures of human foreskin keratinocytes (HFKs) with stable expression of HPV16 E6/E7 or a control vector. After integration of the two data sets, we identified 51 differentially expressed cellular miRs associated with the modulation of 1,456 potential target mRNAs in HPV16 E6/E7-expressing HFKs. We discovered that the degree of differential miR expression in HFKs expressing HPV16 E6/E7 was not necessarily predictive of the number of corresponding mRNA targets or the potential impact on gene expression. Additional analyses of the identified miR-mRNA pairs suggest modulation of specific biological activities and biochemical pathways. Overall, our study supports the model that perturbation of cellular miR expression by HPV16 E6/E7 importantly contributes to the rewiring of cellular regulatory circuits by the high-risk HPV E6 and E7 proteins that contribute to oncogenic transformation.
High-risk human papillomaviruses (HPVs) are the causative agents of almost all cervical cancers and many other cancers, including anal, vaginal, vulvar, penile, and oropharyngeal cancers. Despite the availability of efficacious HPV vaccines, it is critical to determine how HPVs cause cancer, as many people remain unvaccinated and the vaccine does not prevent cancer development in individuals who are already infected. Two HPV proteins, E6 and E7, are the major drivers of cancer development, and much remains to be learned about how the expression of these viral proteins reprograms infected cells, ultimately resulting in cancer development. Small, noncoding human RNAs, termed microRNAs (miRs), regulate gene expression and have been implicated in almost all human cancers, including HPV-associated cancers. Our study provides a comprehensive analysis of how E6 and E7 alter the expression of human miRs and how this potentially impacts cellular gene expression, which may contribute to cancer development.
E6和E7蛋白是高危型人乳头瘤病毒（HPV）编码的主要致癌驱动因子。虽然这些蛋白转化活性的许多方面已得到广泛研究，但较少有研究探讨HPV E6/E7表达如何影响细胞非编码RNA的表达。我们研究的目的是调查HPV16 E6/E7对细胞微小RNA（miR）水平的调节作用，并确定其对细胞基因表达的潜在影响。我们对稳定表达HPV16 E6/E7或对照载体的人包皮角质形成细胞（HFK）原代未分化培养物中的小细胞RNA和大细胞RNA进行了深度测序。整合这两个数据集后，我们在表达HPV16 E6/E7的HFK中鉴定出51种差异表达的细胞miR，它们与1456个潜在靶mRNA的调节相关。我们发现，在表达HPV16 E6/E7的HFK中，miR差异表达的程度不一定能预测相应mRNA靶标的数量或对基因表达的潜在影响。对鉴定出的miR-mRNA对的进一步分析表明，特定的生物学活性和生化途径受到了调节。总体而言，我们的研究支持这样一种模型，即HPV16 E6/E7对细胞miR表达的干扰在很大程度上导致了高危型HPV E6和E7蛋白对细胞调节回路的重新布线，从而促进致癌转化。
高危型人乳头瘤病毒（HPV）是几乎所有宫颈癌以及许多其他癌症（包括肛门癌、阴道癌、外阴癌、阴茎癌和口咽癌）的病原体。尽管有有效的HPV疫苗，但确定HPV如何致癌至关重要，因为许多人仍未接种疫苗，而且该疫苗不能预防已感染个体的癌症发展。两种HPV蛋白E6和E7是癌症发展的主要驱动因子，关于这些病毒蛋白的表达如何对感染细胞进行重新编程，最终导致癌症发展，仍有许多有待了解的地方。小型非编码人类RNA，即微小RNA（miR），可调节基因表达，几乎涉及所有人类癌症，包括与HPV相关的癌症。我们的研究全面分析了E6和E7如何改变人类miR的表达，以及这如何潜在地影响细胞基因表达，而这可能有助于癌症发展。
