甲基化敏感性高分辨率熔解曲线检测结直肠癌组织及外周血MGMT基因甲基化  

Detection of MGMT methylation levels in tumour tissue and peripheral blood of colorectal cancer patients by methylation-sensitive high-resolution melting analysis

作　　者：卫旭宇[1] 张宏莲[2] 高晓云[1] 王萍[2] 毛红菊[2] 金庆辉[2] 赵建龙[2] 马天乐[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院消化内科,上海200025 [2]中国科学院上海微系统与信息技术研究所,上海200050

出　　处：《临床检验杂志》2014年第11期806-809,816,共5页Chinese Journal of Clinical Laboratory Science

基　　金：国家高技术研究发展计划(973计划;2012CB933303);上海市科委项目(12441902600;11nm0505800;12nm0503702)

摘　　要：目的用甲基化敏感性高分辨率熔解曲线法（MS-HRM）定量检测结直肠癌组织及外周血中MGMT基因甲基化水平,并结合结直肠癌病理参数分析探讨MGMT基因甲基化的临床意义。方法以33例结直肠癌组织和配对的正常黏膜组织以及其中16例患者的外周血为研究对象,用MS-HRM进行MGMT基因甲基化检测,分析MGMT基因甲基化与结直肠癌临床病理特征的关系和MS-HRM的方法特点。结果 MS-HRM从33例结直肠癌患者的癌组织中检测到12例（36.4%）MGMT基因启动子不同程度的甲基化（甲基化〈5%1例,5%-10%3例,10%-25%3例,25%-50%2例,50%-75%3例）,对应的正常结直肠组织中仅检测到甲基化〈5%1例（3.0%）。16例外周血中检测到甲基化小于〈5%3例（18.8%）。MGMT基因甲基化与患者的年龄、性别、肿瘤部位无相关性（P〉0.05）,但与结直肠癌的病理分期和分化程度的相关性有统计学意义（P〈0.05）。结论 MS-HRM可定量检测结直肠癌MGMT甲基化水平。MGMT基因甲基化与结直肠癌的进展评估有关。Objective To quantitatively detect the O6-methylguanine-DNA-methyltransferase（MGMT） methylation levels in tumor tis- sue and peripheral blood of colorectal cancer patients by methylation-sensitive high-resolution melting（MS-HRM） analysis, and evalu- ate its clinical significance based on pathological parameters. Methods The MGMT methylation levels in tumor tissues and corre- sponding normal mucous membrane tissues from 33 patients with colorectal canser and in peripheral blood samples from 16 patients were determined by MS-HRM. The relationship between MGMT methylation levels and the clinical-pathological features of colorectal cancer were analyzed and the detection performance of the MS-I-IRM was evaluated. Results The methylation of MGMT promoter re- gion was detected in 36.4% （12/33） of tumor tissue samples, including 1 with 〈 5% of methylation, 3 with 5% -10%, 3 with 10% - 25% , 2 with 25% -50%, and 3 with 50% -75% , while 3.0% （1/33） of normal mucous membrane tissue and 18.8% （3/16） of pe- ripheral blood samples with 〈 5% of methylation. The MGMT methylation levels were not related to patients＇ age, gender and tumor lo- cation（ P 〉 0.05 ）, but significantly related to the pathological stage and differentiation of colorectal cancer（ P 〈0.05 ）. Conclusion The MS-HRM method may be used to quantitatively detect the MGMT methylation levels in the patients with colorectal cancer, and the MGMT methylation may be related to the progression of colorectal cancer.

关 键 词：O6-甲基鸟嘌呤-DNA-甲基转移酶 结直肠癌 甲基化 甲基化敏感性高分辨率熔解曲线 

分 类 号：R446[医药卫生—诊断学]