High-Definition DNA Methylation Profiles from Breast and Ovarian Carcinoma Cell Lines with Differing Doxorubicin Resistance
High-Definition DNA Methylation Profiles from Breast and Ovarian Carcinoma Cell Lines with Differing Doxorubicin Resistance
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DOI：
10.1371/journal.pone.0011002
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发表时间：
2010-06-08
期刊：
影响因子：
3.7
通讯作者：
Hoheisel, Joerg D.
中科院分区：
文献类型：
--
作者：
Boettcher, Michael;Kischkel, Frank;Hoheisel, Joerg D.
获得性耐药是阻碍癌症有效化疗的常见障碍。在过去的几十年里，异常DNA甲基化对耐药肿瘤细胞发展的贡献越来越受到关注。因此，本研究的目的是表征由化疗药物阿霉素治疗肿瘤细胞引起的DNA甲基化变化。研究人员分析了与28个基因相关的CpG岛（cgi）的DNA甲基化水平，这些基因的表达水平先前已被证明有助于抵抗DNA双链断裂诱导药物或不同癌症类型的肿瘤进展。确定了高清晰度DNA甲基化谱，其中包括从800个CpG位点到选定基因转录起始位点周围的cgi的甲基化水平。为了研究CGI甲基化对相关基因表达的影响，我们通过qRT-PCR检测了相关基因的mRNA水平。结果表明，该方法适用于提供高度精确的甲基化图谱，可与克隆测序获得的图谱相媲美，克隆测序是高清DNA甲基化研究的金标准。在对诱导双链断裂药物阿霉素获得性耐药的乳腺癌细胞中，检测到与13个基因相关的CGIs特异性胞嘧啶甲基化的变化。此外，从获得性阿霉素耐药的乳腺癌和卵巢癌细胞系中获得的甲基化谱之间存在相似性。分析的基因子集的表达水平被证明与分析的cgi的甲基化水平有关。我们的研究结果为获得性阿霉素耐药性的两个独立模型系统提供了详细的DNA甲基化信息，并表明在暴露于药物后，两个系统都发生了类似的甲基化变化。
Acquired drug resistance represents a frequent obstacle which hampers efficient chemotherapy of cancers. The contribution of aberrant DNA methylation to the development of drug resistant tumor cells has gained increasing attention over the past decades. Hence, the objective of the presented study was to characterize DNA methylation changes which arise from treatment of tumor cells with the chemotherapeutic drug doxorubicin. DNA methylation levels from CpG islands (CGIs) linked to twenty-eight genes, whose expression levels had previously been shown to contribute to resistance against DNA double strand break inducing drugs or tumor progression in different cancer types were analyzed. High-definition DNA methylation profiles which consisted of methylation levels from 800 CpG sites mapping to CGIs around the transcription start sites of the selected genes were determined. In order to investigate the influence of CGI methylation on the expression of associated genes, their mRNA levels were investigated via qRT-PCR. It was shown that the employed method is suitable for providing highly accurate methylation profiles, comparable to those obtained via clone sequencing, the gold standard for high-definition DNA methylation studies. In breast carcinoma cells with acquired resistance against the double strand break inducing drug doxorubicin, changes in methylation of specific cytosines from CGIs linked to thirteen genes were detected. Moreover, similarities between methylation profiles obtained from breast and ovarian carcinoma cell lines with acquired doxorubicin resistance were found. The expression levels of a subset of analyzed genes were shown to be linked to the methylation levels of the analyzed CGIs. Our results provide detailed DNA methylation information from two separate model systems for acquired doxorubicin resistance and suggest the occurrence of similar methylation changes in both systems upon exposure to the drug.