p16, MGMT, RARbeta2, CLDN3, CRBP and MT1G gene methylation in esophageal squamous cell carcinoma and its precursor lesions.
p16, MGMT, RARbeta2, CLDN3, CRBP and MT1G gene methylation in esophageal squamous cell carcinoma and its precursor lesions.
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DOI：
10.3892/or.15.6.1591
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发表时间：
2006-06
期刊：
影响因子：
4.2
通讯作者：
M. Roth;C. Abnet;N. Hu;Quan-hong Wang;Weng-Qiang Wei;L. Green;Mary D'Alelio;Y. Qiao;S. Dawsey;P. Taylor;K. Woodson
中科院分区：
文献类型：
--
作者：
M. Roth;C. Abnet;N. Hu;Quan-hong Wang;Weng-Qiang Wei;L. Green;Mary D'Alelio;Y. Qiao;S. Dawsey;P. Taylor;K. Woodson
食管鳞状细胞癌是一种常见的恶性肿瘤，预后极差。需要新的方法来筛查高危人群，并早期识别可治愈的肿瘤和前驱病变。分子标记可能在这种筛选工作中是有用的。本研究旨在确定ESCC患者中p16、MGMT、RAR β 2、CLDN 3、CRBP和MT 1G基因甲基化的患病率，以评估癌前病变范围内基因甲基化的变化，并评估在利用球囊细胞学采样器收集的冷冻食管细胞进行初步筛查试验中使用基因甲基化的可行性。样本来自中国中北部的高风险受试者。这些样本包括来自6个完全包埋切除标本的11个组织学正常粘膜病灶、8个低度鳞状不典型增生病灶、7个高度鳞状不典型增生病灶和13个ESCC病灶;来自6个无疾病组织学证据的个体的内窥镜活检;以及来自12个无症状受试者的冷冻食管球囊样本。使用基于Taqman化学的实时甲基化特异性PCR（qMS-PCR）确定每个基因的启动子CpG位点特异性超甲基化状态。6例ESCC患者中，5例至少有一个基因发生甲基化。对于大多数基因，甲基化在肿瘤进展期间发生频率增加，在低度和高度异型增生之间发现最大的增加。在相同组织学分级的不同病灶之间，甚至在个体患者中，甲基化模式存在相当大的差异，但16/20（80%）的高度异型增生和癌灶具有>或= 2个甲基化基因，而17/19（89%）的正常和低度异型增生病灶具有<2个甲基化基因。这些基因很少甲基化，在组织学正常的粘膜患者或没有食管鳞癌。基因甲基化是常见的，很容易检测到的冷冻食管细胞收集气球细胞学采样器。我们的数据表明，p16、MGMT、RAR β 2、CLDN 3、CRBP和MT 1G的甲基化在ESCC患者的食管粘膜中很常见，并且随着疾病组织学严重程度的增加，在病灶中的患病率往往会增加。来自基因组的甲基化数据可能能够识别具有高级别病变的患者。球囊细胞学检查可能能够有效地筛查食管长度的异常甲基化细胞亚群，并可能在ESCC及其前驱病变的初步筛查试验中有用。
Esophageal squamous cell carcinoma (ESCC) is a common cancer with a very poor prognosis. New methods are needed to screen high-risk populations and identify curable tumors and precursor lesions early. Molecular markers may be useful in such screening efforts. This study was designed to determine the prevalence of p16, MGMT, RARbeta2, CLDN3, CRBP and MT1G gene methylation in patients with ESCC to evaluate the variation of gene methylation across a spectrum of preneoplastic lesions, and assess the feasibility of using gene methylation in a primary screening test utilizing frozen esophageal cells collected by balloon cytology samplers. Samples were obtained from high-risk subjects from north central China. These samples included 11 foci of histologically normal mucosa, 8 foci of low-grade squamous dysplasia, 7 foci of high-grade squamous dysplasia, and 13 foci of ESCC from 6 fully embedded resection specimens; endoscopic biopsies from 6 individuals with no histological evidence of disease; and frozen esophageal balloon samples from 12 asymptomatic subjects. Promoter CpG site-specific hypermethylation status was determined for each gene using real-time methylation-specific PCR (qMS-PCR) based on Taqman chemistry. Of the 6 ESCC patients, 5 showed methylation of at least one gene. For most genes, methylation occurred with increasing frequency during neoplastic progression, with the largest increase found between low- and high-grade dysplasia. There was considerable variation in methylation patterns among different foci of the same histological grade, even within individual patients, but 16/20 (80%) of high-grade dysplastic and cancer foci had >or= 2 methylated genes, while 17/19 (89%) of normal and low-grade dysplastic foci had <2 methylated genes. These genes were rarely methylated in histologically normal mucosa from patients with or without ESCC. Gene methylation was common and easily detectable in the frozen esophageal cells collected by balloon cytology samplers. Our data suggest that methylation of p16, MGMT, RARbeta2, CLDN3, CRBP, and MT1G is common in the esophageal mucosa of patients with ESCC in this high-risk population, and tends to increase in prevalence in foci with increasing histological severity of disease. Methylation data from panels of genes may be able to identify patients with high-grade lesions. Balloon cytology may be able to screen the length of the esophagus effectively for a subset of cells with abnormal methylation, and may be useful in a primary screening test for ESCC and its precursor lesions.