Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands.
Genes Chromosomes Cancer. 2013 Jan;52(1):56-68. doi: 10.1002/gcc.22006. Epub 2012 Sep 14.
Cervical cancer results from persistent infection with high-risk human papillomavirus (hrHPV). Common genetic aberrations in cervical (pre)cancers encompass large genomic regions with numerous genes, hampering identification of driver genes. This study aimed to identify genes functionally involved in HPV-mediated transformation by analysis of focal aberrations (<3 Mb) in high-grade cervical intraepithelial neoplasia (hgCIN). Focal chromosomal aberrations were determined in high-resolution array comparative genomic hybridization data of 60 hgCIN. Genes located within focal aberrations were validated using 2 external gene expression datasets or qRT-PCR. Functional roles of candidate genes EYA2 (20q13) and hsa-miR-375 (2q35) were studied by siRNA-mediated knock-down and overexpression, respectively, in hrHPV-containing cell lines. We identified 74 focal aberrations encoding 305 genes. Concurrent altered expression in hgCIN and/or cervical carcinomas compared with normal cervical samples was shown for ATP13A3, HES1, OPA1, HRASLS, EYA2, ZMYND8, APOBEC2, and NCR2. Gene silencing of EYA2 significantly reduced viability, migratory capacity, and anchorage-independent growth of HPV16-transformed keratinocytes. For hsa-miR-375, a direct correlation between a (focal) loss and significantly reduced expression was found. Downregulation of hsa-miR-375 expression was confirmed in an independent series of cervical tissues. Ectopic expression of hsa-miR-375 in 2 cervical carcinoma cell lines reduced cellular viability. Our data provide a proof of concept that chromosomal aberrations are actively contributing to HPV-induced carcinogenesis and identify EYA2 and hsa-miR-375 as oncogene and tumor suppressor gene, respectively.
宫颈癌是由高危型人乳头瘤病毒（hrHPV）持续感染引起的。在宫颈癌（前）癌中常见的遗传异常包括大片段基因组区域和众多基因，这阻碍了驱动基因的鉴定。本研究旨在通过分析高级别宫颈上皮内瘤变（hgCIN）中的局灶性染色体异常（<3 Mb），鉴定参与 HPV 介导转化的功能相关基因。使用高分辨率阵列比较基因组杂交数据，在 60 例 hgCIN 中确定局灶性染色体异常。使用 2 个外部基因表达数据集或 qRT-PCR 验证位于局灶性异常内的基因。通过 siRNA 介导的敲低和过表达，分别研究候选基因 EYA2（20q13）和 hsa-miR-375（2q35）在含有 hrHPV 的细胞系中的功能作用。我们鉴定了 74 个编码 305 个基因的局灶性异常。与正常宫颈样本相比，在 hgCIN 和/或宫颈癌中，ATP13A3、HES1、OPA1、HRASLS、EYA2、ZMYND8、APOBEC2 和 NCR2 的表达发生了改变。EYA2 的基因沉默显著降低了 HPV16 转化的角质形成细胞的活力、迁移能力和锚定非依赖性生长。对于 hsa-miR-375，发现了一个（局灶性）缺失和显著降低表达之间的直接相关性。在独立的宫颈组织系列中，证实了 hsa-miR-375 的下调。在 2 种宫颈癌细胞系中过表达 hsa-miR-375 降低了细胞活力。我们的数据提供了一个概念验证，即染色体异常积极参与 HPV 诱导的致癌作用，并分别鉴定 EYA2 和 hsa-miR-375 为癌基因和肿瘤抑制基因。
