Promoter methylation of P16, RARbeta, E-cadherin, cyclin A1 and cytoglobin in oral cancer: quantitative evaluation using pyrosequencing.
Promoter methylation of P16, RARbeta, E-cadherin, cyclin A1 and cytoglobin in oral cancer: quantitative evaluation using pyrosequencing.
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口腔癌中p16，rarbeta，e-钙粘蛋白，细胞周期蛋白A1和细胞神经蛋白的启动子甲基化：使用焦磷酸测序进行定量评估。
DOI：
10.1038/sj.bjc.6602972
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发表时间：
2006-02-27
影响因子：
8.8
通讯作者：
中科院分区：
文献类型：
--
作者：
癌组织的甲基化谱已经确定这种机制是致癌作用的重要组成部分。通过启动子甲基化的肿瘤抑制基因的表观遗传沉默已经通过各种手段进行了研究，其中最新的是焦磷酸测序。我们研究了口腔鳞状细胞癌患者的定量甲基化状态。新鲜肿瘤组织和正常对照组织的切除边缘，从79个连续的患者进行切除的口腔鳞状细胞癌。提取DNA并进行亚硫酸氢盐处理。设计PCR引物扩增p16、RARβ、E-cadherin、cytoglobin和cyclinA 1基因启动子的75-200 bp区域。通过焦磷酸测序确定每个基因4-5个CpG位点的甲基化状态。在肿瘤标本中发现28%的p16、73%的RARβ、42%的E-钙粘蛋白、65%的细胞珠蛋白和53%的细胞周期蛋白A1基因启动子的显著CpG甲基化。与正常组织相比，肿瘤中p16（P=0.048）、细胞珠蛋白（P=0.002）和细胞周期蛋白A1（P=0.001）的启动子甲基化显著升高，但RARβ（P=0.088）或E-cadherin（P=0.347）的启动子甲基化没有升高。在该肿瘤系列中证明了一致的甲基化（P=0.03）。除RARβ外，所有基因的单个CpG位点甲基化程度均存在显著差异，这些差异在肿瘤样本之间重现。细胞周期蛋白A1启动子甲基化与组织学分级呈相反趋势。使用焦磷酸测序的启动子甲基化分析揭示了来自几个CpG位点的有价值的定量数据。与甲基化特异性PCR产生的定性数据相反，我们的数据表明p16启动子甲基化具有高度的肿瘤特异性。还观察到显著的肿瘤特异性细胞周期蛋白A1启动子甲基化。细胞珠蛋白是上呼吸消化道鳞癌中高度甲基化的一个新的候选抑癌基因。
Methylation profiling of cancer tissues has identified this mechanism as an important component of carcinogenesis. Epigenetic silencing of tumour suppressor genes through promoter methylation has been investigated by a variety of means, the most recent of which is pyrosequencing. We have investigated quantitative methylation status in oral squamous cell carcinoma patients. Fresh tumour tissue and normal control tissue from resection margin was obtained from 79 consecutive patients undergoing resection of oral squamous cell carcinoma. DNA was extracted and bisulphite treated. PCR primers were designed to amplify 75–200 bp regions of the CpG rich gene promoters of p16, RARβ, E-cadherin, cytoglobin and cyclinA1. Methylation status of 4-5 CpG sites per gene was determined by pyrosequencing. Significant CpG methylation of gene promoters within tumour specimens was found in 28% for p16, 73% for RARβ, 42% for E-cadherin, 65% for cytoglobin and 53% for cyclinA1. Promoter methylation was significantly elevated in tumours compared to normal tissue for p16 (P=0.048), cytoglobin (P=0.002) and cyclin A1 (P=0.001) but not in RARβ (P=0.088) or E-cadherin (P=0.347). Concordant methylation was demonstrated in this tumour series (P=0.03). Significant differences in degree of methylation of individual CpG sites were noted for all genes except RARβ and these differences were in a characteristic pattern that was reproduced between tumour samples. Cyclin A1 promoter methylation showed an inverse trend with histological grade. Promoter methylation analysis using pyrosequencing reveals valuable quantitative data from several CpG sites. In contrast to qualitative data generated from methylation specific PCR, our data demonstrated p16 promoter methylation in a highly tumour specific pattern. Significant tumour specific methylation of cyclin A1 promoter was also seen. Cytoglobin is a novel candidate tumour suppressor gene highly methylated in upper aero-digestive tract squamous cancer.
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DOI：
10.1073/pnas.93.18.9821
发表时间：
1996-09-03
影响因子：
11.1
作者：
Herman, JG;Graff, JR;Baylin, SB
通讯作者：
Baylin, SB
影响因子：
5.3
作者：
Langan, JE;Cole, CG;Risk, JM
通讯作者：
Risk, JM
影响因子：
3.5
作者：
Huang, THM;Perry, MR;Laux, DE
通讯作者：
Laux, DE
影响因子：
2.7
作者：
Colella, S;Shen, L;Krahe, R
通讯作者：
Krahe, R
影响因子：
8
作者：
Coombes, MM;Briggs, KL;Dent, SYR
通讯作者：
Dent, SYR