巢式甲基化特异性PCR检测p16基因的研究

Detection of hypermethylation of p16 gene from colorectal cancer patient

目的检测p16基因启动子区异常甲基化发生情况,探讨p16基因异常甲基化作为结直肠癌临床辅助诊断分子生物学标志物的可能性。方法应用巢式甲基化特异性PCR技术(Nested-MSP)及甲基化特异性PCR(MSP),检测了结直肠癌患者肿瘤组织中p16基因的异常甲基化情况;比较MSP及Nested-MSP两方法的灵敏度。结果应用MSP方法,Dukes A、B期病人和Dukes C、D期病人p16启动子区甲基化率分别为23.8%和59.4%;应用巢式-MSP方法,甲基化率上升至52.4%和84.4%。结论巢式-MSP的灵敏度明显高于普通的MSP技术,分析患者DNA的p16基因异常甲基化有可能成为辅助结直肠癌诊断的有效方法之一。

Objective To detect hypermethylation of p16 gene in tissue DNA from patients with colorectal cancer,and to assess its potential as a malignant marker. Methods Using methylation specific PCR （MSP） and nested methylation specific PCR （Nested-MSP） ,the status of methylation of the p16 was investigated in tumor tissues DNA from 53 colorectal cancer patients. Results The hypermethylation of the p16 was present in 23.8% in Dukes stages A and B tumors,59.4% in the stages of C and D tumors by MSP; the hypermethylation of the p16 was present in 52.4% in Dukes stages A and B tumors, 84.4% in the stages of C and D tumors by Nested-MSP. Conclusion The result indicates that the sensitivity of MSP is higher than that of Nested-MSP significantly, and the hypermethylation of the p16 may be a useful marker in the auxiliary diagnosis of colorectal cancers.