MSRE-qPCR技术分析多基因DNA甲基化对肝细胞癌的诊断价值  

The methylation analysis of multiple genes using methylation-sensitive restriction enzymes-based quantitative PCR for the detection of hepatocellular carcinoma

作　　者：邱东民[1,2] 余坚[3] 黄朝晖[2] 

机构地区：[1]无锡市卫生高等职业技术学校,江苏无锡214021 [2]苏州大学附属第四医院肿瘤研究所,江苏无锡214062 [3]上海市肿瘤研究所癌基因与癌相关基因国家重点实验室,上海200032

出　　处：《中国癌症杂志》2011年第8期615-620,共6页China Oncology

基　　金：江苏省自然科学基金资助项目(No:BK2008114)

摘　　要：背景与目的:DNA甲基化是潜在的肿瘤标志物。本研究运用自行建立的甲基化敏感性限制性内切酶-定量PCR(methylation-sensitive restriction enzymes-based quantitative PCR,MSRE-qPCR)方法检测肝细胞癌(hepatocellutar carcinoma,HCC)组织中多个基因的DNA甲基化状态,筛选可用于HCC诊断的DNA甲基化标志物组合。方法:以47对HCC患者癌组织和癌旁组织、8例正常人肝组织为材料,运用MSRE-qPCR方法检测这102例肝组织中APC、GSTP1、RASSF1A、p16、SFRP1、RUNX3、Hint1、SOCS1和HIC-1基因的启动子甲基化状态。结果:APC、GSTP1、RASSF1A、p16、SFRP1、RUNX3基因在肿瘤组织中的甲基化率分别为70.2%、70.2%、63.8%、29.8%、44.7%和36.2%,均显著高于对应癌旁组织(P均<0.05);而Hint1、SOCS1和HIC-1基因甲基化水平在HCC肿瘤和癌旁组织间无显著差异。联合检测APC、GSTP1、RASSF1A和SFRP1 4个靶点,可检出所有HCC病例。这些基因的甲基化水平与患者肿瘤大小、分化、包膜及HBV感染等临床病理参数均无显著相关性。结论:联合检测APC、GSTP1、RASSF1A和SFRP1的DNA甲基化状态对于HCC风险评估和分子诊断具有重要价值。Background and purpose： DNA methylation is a promising biomarker for cancer. This study was aimed to investigate the methylation status of multiple genes using methylation-sensitive restriction enzymes-based quantitative PCR （MSRE-qPCR） method in hepatocellular carcinoma （HCC） and identify a combination of methyaltion markers for the diagnosis of HCC. Methods： One hundred and two live tissues, including 8 normal live tissues, 47 paired HCC and non-cancerous tissues, were collected. The methylation status ofAPC, GSTP1, RASSF1A,p16, SFRP1, RUNX3, SOCS1, Hintl and HIC-1 genes in these live tissues were determined using MSRE-qPCR method. Results： The methylation rates ofAPC, GSTP1, RASSF1A, p16, SFRP1 and RUNX3 genes in HCC were 70.2%, 70.2%, 63.8%, 29.8%, 44.7% and 36.2%, respectively, which were significantly higher than those in adjacent non-cancerous tissues （P〈0.05）. No significant difference was found about the methylation rate of SOCS1, Hintl and HIC-1 genes between HCC and adjacent tissues. The combination analysis ofAPC, GSTP1, RASSF1A and SFRP1 could detect all these 47 HCC cases. The methylation status of these genes was not associated with tumor size, tumor differentiation, tumor capsule and HBV infection. Conclusion： DNA methylation analysis ofAPC, GSTPI, RASSFIA and SFRP1 may be a promising method for the risk assessment and early diagnosis of HCC.

关 键 词：DNA甲基化 限制性内切酶 肝细胞癌 诊断 

分 类 号：R735.7[医药卫生—肿瘤]