巢式甲基化特异性PCR 检测食管癌病人血清中p16 基因启动子区过甲基化

 目的 检测食管鳞状细胞癌（squamous cell carcinoma，SCC）患者外周血血清中p16基因启动子区的甲基化状态，探讨p16基因启动子的过甲基化在食管鳞状细胞癌筛查及早期诊断中的意义。方法 利用巢式甲基化特异性PCR（nMSP）法检测食管鳞状细胞癌患者外周血血清与正常人血清中p16基因启动子的甲基化状态，并与普通甲基化特异性PCR（MSP）法进行了比较。结果 56例SCC血清样品中nMSP法发现34例p16基因启动子的过甲基化，MSP法只检出15例，而22例正常人血清中都未检测到p16基因启动子的过甲基化。测序结果进一步验证了方法的可靠性。结论 利用巢式MSP（nMSP）法检测外周血血清中p16基因启动子的甲基化，可为食管癌的筛查、早期诊断及预后判断提供有价值的信息。

Detection of Promoter Hypermethylation in the Serum of Esophageal Squamous Cell Carcinoma Patients by Nested Methylation-Specific-Polymerase Chain Reaction

Objective This study was designed to detect the methylation status of the promoter region of p16gene in DNA extracted serum from esophageal squamous cell carcinoma (SCC) ,and evaluate the role of p16gene promoter hypermethylation in esophageal SCC screening and early diagnosis.Methods Nes- ted methylation-specific PCR (nMSP) was used to detect p16promoter hypermethylation in serum DNA from56esophageal squamous cell carcinoma (SCC) and control serum samples from22healthy individu- als.Compared methylation-specific PCR(MSP), nMSP was more sensitive.Results Aberrant promoter rmethylation of the p16gene was found in34of56serum samples using nMSP ,and only15cases was detected using MSP. No aberrant promoter methylation was detected in the peripheral serum of the healthy individuals.Conclusion The detection of p16gene promoter hypermethylation in the serum of e- sophageal SCC patients can give the useful information for tumor early diagnosis , follow-up study of e- sophageal SCC patients. nMSP is a simple,sensitive ,and specific method for rapid analysis of the pro- moter methylation status of many genes.