Yablonska Svitlana, Hoskins Elizabeth E, Wells Susanne I, Khan Saleem A
Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
Microrna. 2013;2(1):2-13. doi: 10.2174/2211536611302010002.
Human papillomaviruses (HPVs) are associated with the pathogenesis of a variety of human cancers, including cervical and oropharyngeal cancers. The HPV E6 and E7 oncogenes are usually expressed to high levels in these cancers. Previous studies have shown dysregulation of cellular microRNAs (miRNAs) in HPV-positive cell lines and cancer tissues and recent studies have identified a few miRNAs whose levels are altered in the presence of the viral E6 and E7 proteins. In order to identify all the cellular miRNAs whose expression may be affected by these oncoproteins, we carried out microarray analysis using human foreskin keratinocytes (HFKs) expressing either or both of these two proteins. These studies showed that 90 and 60 miRNAs were dysregulated in the presence of the E6 or the E7 protein, respectively. Of these, 43 miRNAs were similarly affected in HFK-E6 and/or HFK-E7 when compared to control cells. The joint expression of E6 and E7 proteins in HFKs caused changes in the levels of 64 miRNAs, of which 24 were similarly affected in HFK-E6 and/or HFK-E7 cells relative to controls. The microarray experiments were validated by quantitative real-time RT-PCR analysis of several differentially expressed miRNAs. Several miRNAs dysregulated by the E6 and/or E7 proteins are known to be altered in a variety of human cancers. Furthermore, previously known cellular targets of these miRNAs are involved in processes such as cell cycle regulation, apoptosis, cell-cell adhesion, cell mobility and proliferation, and alterations in their levels may contribute to HPV-associated carcinogenesis. Taken together, the results of our studies suggest that high risk HPV E6 and E7 proteins share the ability to regulate a subset of cellular miRNAs.
人乳头瘤病毒（HPV）与多种人类癌症的发病机制相关，包括宫颈癌和口咽癌。HPV E6和E7癌基因通常在这些癌症中高表达。先前的研究表明，HPV阳性细胞系和癌组织中细胞微小RNA（miRNA）存在失调，最近的研究已经鉴定出一些在病毒E6和E7蛋白存在时其水平发生改变的miRNA。为了鉴定所有其表达可能受这些癌蛋白影响的细胞miRNA，我们使用表达这两种蛋白之一或两者的人包皮角质形成细胞（HFK）进行了微阵列分析。这些研究表明，分别有90个和60个miRNA在E6或E7蛋白存在时失调。其中，与对照细胞相比，43个miRNA在HFK-E6和/或HFK-E7中受到类似影响。HFK中E6和E7蛋白的联合表达导致64个miRNA水平发生变化，其中24个在HFK-E6和/或HFK-E7细胞中相对于对照受到类似影响。通过对几种差异表达的miRNA进行定量实时RT-PCR分析验证了微阵列实验。已知一些被E6和/或E7蛋白失调的miRNA在多种人类癌症中发生改变。此外，这些miRNA先前已知的细胞靶标参与细胞周期调控、细胞凋亡、细胞间粘附、细胞迁移和增殖等过程，其水平的改变可能有助于HPV相关的致癌作用。综上所述，我们的研究结果表明，高危型HPV E6和E7蛋白具有共同调节一部分细胞miRNA的能力。
