应用Methylight法检测胃癌病人p16基因的甲基化状态

Detecting the Methylation Status of p16 Gene in Gastric carcinomas patients by Methylight Method

目的探讨用Methylight法检测p16基因启动子区甲基化的可行性以及在肿瘤分子诊断中的应用价值。方法从45例原发性胃癌患者的石蜡包埋肿瘤组织中提取基因组DNA,通过焦亚硫酸钠修饰将未甲基化的胞嘧啶转化为尿嘧啶。通过115bp的Methylight法和经典的MSP法检测p16基因的甲基化状态。结果以甲基化特异性PCR(Methylation-specific PCR,MSP)MSP法做为金标准,我们发现Methylight法检测p16基因甲基化的灵敏度、特异性和符合率分别为88.2%、92.9%和91.1%。结论 Methylight法是一种可靠、实用的检测p16甲基化的方法,可用于肿瘤的分子学诊断。

Objective To investigate the promoter methylation of p16 gene by a 115-bp Methylight assay and its possible application in molecular diagnosis of tumor.Methods Genomic DNA of 45 primary gastric carcinomas patients were extracted from paraffin-embedded tumor tissue and modified with sodium bisulfite to convert the unmethylated cytosines to uridines.The methylation status of p16 gene was determined by a 115-bp Methylight assay and Classic Methylation-specific PCR(MSP).Results Using the results of MSP as a golden standard,we found the sensitivity,specificity and coincidence for Methylight assay to be 88.2%,92.9%and 91.1%.Conclusion The Methylight assay may be an eligible and practical method for the detection of methylated-p16 biomarker in molecular diagnosis of tumor.