Identification and Validation of DNA Methylation Markers to Predict Lymph Node Metastasis of Esophageal Squamous Cell Carcinomas
Identification and Validation of DNA Methylation Markers to Predict Lymph Node Metastasis of Esophageal Squamous Cell Carcinomas
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DOI：
10.1245/s10434-010-1393-5
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发表时间：
2011-04-01
影响因子：
3.7
通讯作者：
Ushijima, Toshikazu
中科院分区：
文献类型：
--
作者：
Gyobu, Ken;Yamashita, Satoshi;Ushijima, Toshikazu
食管鳞状细胞癌（ESCC）患者是否存在淋巴结转移是决定治疗策略的关键因素。然而，目前还没有评估淋巴结转移的分子标志物。本研究将96例食管鳞癌分为筛选组（n = 48）和验证组（n = 48），研究CpG岛甲基化与食管鳞癌淋巴结转移的关系。全基因组甲基化分析采用甲基化DNA免疫沉淀-CGI微阵列分析。采用定量甲基化特异性PCR（qMSP）方法检测8例有淋巴结转移和4例无淋巴结转移食管鳞癌组织中25个CGIs的甲基化水平。在筛选组中，7个CGIs在有淋巴结转移和无淋巴结转移的ESCC中的甲基化水平存在显著差异（P <0.05），并确定这些CGIs的临界甲基化水平。采用预先固定的临界值对验证集进行分析，并验证PAX6和ENST 00000363328附近的2个CGI的甲基化状态与淋巴结转移的存在相关。使用这2个标志物，预测存在的敏感性为93%，特异性为57%。此外，2种CGIs的甲基化状态与食管鳞癌的无病生存率显著相关（P = 0.006），与食管鳞癌的淋巴结转移显著相关（P <0.05）。这些CGIs是预测淋巴结转移存在的有希望的标志物。
The presence of lymph node metastasis in esophageal squamous cell carcinoma (ESCC) patients is a critical factor for decision of treatment strategy. However, there have been no molecular markers to assess lymph node metastasis. In this study, we aimed to identify CpG islands (CGIs) whose DNA methylation statuses are associated with the presence of lymph node metastasis.A total of 96 ESCCs were divided into a screening set (n = 48) and a validation set (n = 48). Genome-wide methylation analysis was performed by methylated DNA immunoprecipitation-CGI microarray analysis. Methylation levels were analyzed by quantitative methylation-specific PCR (qMSP).Genome-wide methylation analysis identified 25 CGIs differentially methylated between 8 ESCCs with lymph node metastasis and 4 without. In the screening set, 7 CGIs had significantly different methylation levels (P < 0.05) between the ESCCs with and without lymph node metastasis, and cut-off methylation levels for these CGIs were determined. The validation set was analyzed with the prefixed cut-offs, and methylation statuses of 2 CGIs in the vicinities of PAX6 and ENST00000363328 were validated to be associated with the presence of lymph node metastasis. Using these 2 markers, the presence was predicted with a sensitivity of 93% and specificity of 57%. In addition, the methylation statuses of the 2 CGIs were significantly associated with disease-free survival (P = 0.006).Methylation statuses of these 2 CGIs were significantly associated with the presence of lymph node metastasis of ESCCs. These CGIs are promising markers to predict the presence of lymph node metastases.