Sequential gene promoter methylation during HPV-induced cervical carcinogenesis
Sequential gene promoter methylation during HPV-induced cervical carcinogenesis
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DOI：
10.1038/sj.bjc.6604055
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发表时间：
2007-11-14
影响因子：
8.8
通讯作者：
Steenbergen, R. D. M.
中科院分区：
文献类型：
--
作者：
Henken, F. E.;Wilting, S. M.;Steenbergen, R. D. M.
我们的目的是将宫颈癌中发生的 DNA 甲基化事件与 HPV 诱导的转化的不同阶段联系起来。宫颈癌的甲基化特异性多重连接依赖性探针扩增 (MS-MLPA) 分析显示，所分析的 29 个抑癌基因中有 12 个启动子甲基化，其中 MGMT 甲基化最频繁 (92%)。随后，通过 MS-MLPA 分析了 HPV16/18 转染的角质形成细胞 (n = 11) 的连续阶段，从永生前到锚定独立表型。尽管在永生前的细胞中没有明显的甲基化，但向锚定独立的进展与涉及五个基因的频繁甲基化事件的积累有关，所有这些基因在宫颈癌中也被甲基化。 TP73 和 ESR1 甲基化在早期永生细胞中变得明显，随后在晚期永生细胞中出现 RAR β 和 DAPK1 甲基化。 MGMT 的互补甲基化与锚定独立性相关。对代表致瘤表型的九个宫颈癌细胞系的分析显示，除了这五个基因之外，CADM1、CDH13 和 CHFR 也频繁甲基化。总之，宫颈癌中八个复发性甲基化事件可归属于 HPV 诱导转化的不同阶段。因此，我们的体外模型系统提供了一个有价值的工具，可以进一步从功能上解决宫颈癌中常见的表观遗传改变。
We aimed to link DNA methylation events occurring in cervical carcinomas to distinct stages of HPV-induced transformation. Methylation specific-multiplex ligation-dependent probe amplification (MS-MLPA) analysis of cervical carcinomas revealed promoter methylation of 12 out of 29 tumour suppressor genes analysed, with MGMT being most frequently methylated (92%). Subsequently, consecutive stages of HPV16/18-transfected keratinocytes (n = 11), ranging from pre-immortal to anchorage-independent phenotypes, were analysed by MS-MLPA. Whereas no methylation was evident in pre-immortal cells, progression to anchorage independence was associated with an accumulation of frequent methylation events involving five genes, all of which were also methylated in cervical carcinomas. TP73 and ESR1 methylation became manifest in early immortal cells followed by RAR beta and DAPK1 methylation in late immortal passages. Complementary methylation of MGMT was related to anchorage independence. Analysis of nine cervical cancer cell lines, representing the tumorigenic phenotype, revealed in addition to these five genes frequent methylation of CADM1, CDH13 and CHFR. In conclusion, eight recurrent methylation events in cervical carcinomas could be assigned to different stages of HPV-induced transformation. Hence, our in vitro model system provides a valuable tool to further functionally address the epigenetic alterations that are common in cervical carcinomas.