Quantitative promoter methylation analysis of hepatocellular carcinoma, cirrhotic and normal liver
Quantitative promoter methylation analysis of hepatocellular carcinoma, cirrhotic and normal liver
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DOI：
10.1002/ijc.23433
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发表时间：
2008-06-15
影响因子：
6.4
通讯作者：
Usadel, Henning
中科院分区：
文献类型：
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作者：
Harder, Jan;Opitz, Oliver G.;Usadel, Henning
肝细胞癌是世界范围内最常见的癌症之一。目前对其分子发病机制以及DNA甲基化与疾病发生和发展的相关性知之甚少。然而，一些基因的启动子甲基化已被认为是肝细胞癌的潜在标志。应用定量甲基化特异性聚合酶链式反应技术对34例肝细胞癌、34例配对的非恶性、肝硬变和16例正常肝组织中p16(INK4a)、GSTP1、MGMT、DAP-K和APC基因的甲基化状态进行分析。DNA启动子甲基化频率分别为：p16(INK4a)(76%比24%)、GSTP1(53%比32%)、MGMT(6比12%)、DAP-K(18比100%)和APC(100%比100%)。88%的肝细胞癌组织中存在GSTP1和/或p16(INK4a)启动子甲基化。在正常肝组织中，p16(Ink4a)、GSTP1和MGMT启动子未发生甲基化，DAP-K甲基化的发生率为31%，APC的甲基化发生率为100%。除MGMT外，所有基因甲基化启动子DNA的定量水平在不同组织类型中均有显著差异。我们的结果表明，肿瘤相关基因启动子甲基化是肝癌发生过程中的常见事件。值得注意的是，与非恶性肝硬变相比，p16(INK4a)和GSTP1在肝细胞癌中的启动子甲基化水平和频率更高。这表明这些表观遗传事件可能是一个很好的肝癌标志物。这些数据也证明了在给定样本中对甲基化启动子DNA进行量化的重要性，并使用正常组织作为对照。定量检测GSTP1和p16(INK4a)启动子甲基化状态可作为检测肝细胞癌的分子标志物。(C)2008年Wiley-Liss，Inc.
Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide. Little is known about its molecular pathogenesis and the relevance of DNA methylation for disease initiation and progression. Nevertheless, promoter methylation of some genes has been implicated as potential marker for HCC. Thirty-four HCC, 34 matching non-malignant, cirrhotic livers and 16 normal livers were analyzed for the methylation status of the genes p16(INK4a), GSTP1, MGMT, DAP-K and APC by quantitative methylation-specific PCR. DNA promoter methylation frequencies in HCC and matching non-malignant cirrhotic liver, respectively, were as follows: p16(INK4a) (76% vs. 24%), GSTP1 (53% vs. 32%), MGMT (6 vs. 12%), DAP-K (18 vs. 100%) and APC (100 vs. 100%). GSTP1 and/or p16(INK4a), promoter methylation was observed in 88% of the HCC samples. In normal liver tissue, the p16(INk4a), GSTP1 and MGMT promoter were not methylated DAP-K was methylated in 31% and APC even in 100% of normal liver samples. Quantitative levels of methylated promoter DNA of all genes were significantly different in the various tissue types except for MGMT. Our results suggest that promoter methylation of tumor-associated genes is a common event in hepatocarcinogenesis. Significantly, higher levels and frequencies of promoter methylation in HCC were found for p16(INK4a) and GSTP1 compared to non-malignant cirrhotic liver. This indicates that these epigenetic events may serve as a good marker for HCC. These data also demonstrate the importance of the quantification of methylated promoter DNA within a given sample and the use of normal tissue as controls. Quantitative analyses of methylated GSTP1 and p16(INK4a) promoter may serve as a powerful molecular marker in detecting HCC in biopsies. (C) 2008 Wiley-Liss, Inc.