Focal aberrations indicate EYA2 and hsa‐miR‐375 as oncogene and tumor suppressor in cervical carcinogenesis

Abstract

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.

Clinical implications of (epi)genetic changes in HPV-induced cervical precancerous lesions

“…From these regions candidate driver genes can be extracted by analysis of recurrent focal aberrations and/or expression profiling supplemented with functional analysis. This approach has led to the identification of EYA2 and hsa-mir-375 as novel onco-and tumour suppressor genes, respectively, in cervical cancer 43 . In support of these findings, EYA2 has recently been identified as a target of viral integration and a tumour suppressive function of miR-375 has also been corroborated in other studies 44,45 .…”

Evaluation of Expression of 4 Mirnas in Cytological Samples as an Additional Method of Cervical Cancer Diagnosis

“…Таким обра-зом, было выбрано 18 молекул микроРНК, так назы-ваемых «потенциально маркерных» микроРНК: miR-106 [21,24], miR-1246 [25,26], miR-125b [27][28][29], miR-126 [30][31][32], miR-145 [33,34], miR-146a [35,36], miR-146b [37], miR-155 [38,39], miR-192 [40], miR196b [41], miR-200b [42,43], miR-203 [44,45], miR-20a [46], miR-21 [47,48], miR-31 [49]; miR-34a [27,50]; miR-375 [51,52], miR-99a [53]. Их участие в церви-кальном канцерогенезе было подтверждено как мини-мум в 2 независимых исследованиях.…”

Overexpression of miR-30a in lung adenocarcinoma A549 cell line inhibits migration and invasion via targeting &lt;italic&gt;EYA2&lt;/italic&gt;

“…The allosteric inhibitors of the EYA2 phosphatase had also been reported to inhibit EYA2-mediated cell migration in breast cancer [18]. Moreover, the abnormal expression of EYA2 was also found in other cancers such as cervical cancer [19], pancreatic adenocarcinoma [20], colorectal neoplasia [21], epithelial ovarian cancer [22], and particularly NSCLC [23]. A recent study demonstrated that EYA2 is a direct target of miR-30a in breast cancer [6].…”