Gene expression profiling to identify markers associated with deregulated hTERT in HPV-transformed keratinocytes and cervical cancer
Gene expression profiling to identify markers associated with deregulated hTERT in HPV-transformed keratinocytes and cervical cancer
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DOI：
10.1002/ijc.23210
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发表时间：
2008-02-15
影响因子：
6.4
通讯作者：
Steenbergen, Renske D. M.
中科院分区：
文献类型：
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作者：
de Wilde, Jillian;Wilting, Saskia M.;Steenbergen, Renske D. M.
尽管高危人乳头瘤病毒 (HPV) 感染在宫颈癌的发展中起重要作用，但也涉及附加致癌事件。一个关键事件涉及端粒酶活性的增加，这是由于其催化亚基 hTERT 的表达失调所致。我们之前的微细胞介导的染色体转移研究表明，在HPV16永生化角质形成细胞系FK16A和含有HPV16的宫颈癌细胞系SiHa中引入人类6号染色体会诱导生长停滞，这是由于hTERT mRNA表达和端粒酶活性的抑制所致。在此，该模型用于分析 HPV 转化细胞中与 hTERT 失调相关的表达谱。对 12 个 FK16A/6 号染色体杂交体（其中 4 个内源 hTERT 阴性，8 个内源 hTERT 阳性）进行微阵列表达分析，鉴定出 164 个差异表达基因。通过实时 RT-PCR 验证了 5 个基因选择的差异表达。在这 164 个基因中，有 32 个在 hTERT 失调的其他 HPV 转化细胞中也有差异表达。对于其中编码 AQP3 和 MGP 的 2 个基因，分别通过实时 RT-PCR 和免疫组织化学证实了 hTERT 阳性宫颈癌中表达的改变。此外，与没有hTERT mRNA表达升高的宫颈癌前病变相比，在hTERT mRNA表达升高的高级宫颈癌前病变中，MGP蛋白表达增加的频率明显更高。总之，我们确定了 32 个 hTERT mRNA 表达失调的候选生物标志物，这可能有助于识别进展为浸润性癌风险最高的宫颈癌前病变。 (C) 2007 Wiley-Liss, Inc.
Although high-risk human papillomavirus (HPV) infection plays a major role in the development of cervical cancer, additive oncogenic events are involved as well. One key event involves increased activity of telomerase resulting from a deregulated expression of its catalytic subunit hTERT. Our previous microcell-mediated chromosome transfer studies revealed that introduction of human chromosome 6 in the HPV16-immortalized keratinocyte cell line FK16A and in the HPV16-containing cervical cancer cell line SiHa induced growth arrest, resulting from a repression of hTERT mRNA expression and telomerase activity. Here, this model was used to analyze expression profiles associated with hTERT deregulation in HPV-transformed cells. Microarray expression analysis of 12 FK16A/chromosome 6 hybrids, 4 of which were negative for endogenous hTERT and 8 of which were positive for endogenous hTERT, resulted in the identification of 164 differentially expressed genes. Differential expression of a selection of 5 genes was verified by real-time RT-PCR. Of these 164 genes, 32 were also differentially expressed in other HPV transformed cells with deregulated hTERT. For 2 of these genes, encoding AQP3 and MGP, altered expression in hTERT positive cervical carcinomas was confirmed by real-time RT-PCR and immunohistochemistry, respectively. Moreover, increased MGP protein expression was significantly more frequent in high-grade cervical premalignant lesions with elevated hTERT mRNA expression compared to those without. In summary, we identified 32 candidate biomarkers for deregulated hTERT mRNA expression, which may enable the identification of cervical premalignant lesions that are at highest risk to progress to invasive cancer. (C) 2007 Wiley-Liss, Inc.