甲基化特异性PCR检测肺癌p16基因甲基化及其临床意义

目的:探讨甲基化特异性聚合酶链反应(methylation-specific PCR,MSPCR)在肺癌临床诊断中的应用价值.方法:选取56例肺部疾病住院患者手术切除的病变肺组织和相应的支气管肺泡灌洗液(BALF)及痰标本,其中32例为肺癌,24例为良性肺部疾病.标本经一般处理,PCR扩增后,产物经电泳EB染色,紫外灯下观察.结果:32例肺癌组织标本中,14例(43.8%)在p16基因启动子区域呈现异常甲基化,其中9例(64.3%)在相应的BALF中检测出甲基化存在,5例(35.8%)在相应的痰标本中也检测出甲基化存在.24例良性肺部疾病,其中肺囊肿10例,肺结核14例,手术切除标本和BALF及痰标本中均未检测出p16基因甲基化存在.结论:MSPCR技术对肺癌患者BALF及痰标本中的异常甲基化检测具有高度特异性,是一项有潜力的肺癌早期诊断新技术.

Detection of p16 Gene Methylation by Methylation-specific PCR and Its Clinical Significance in Lung Cancer

Objective: To explore the clinical value of methylation-specific polymerase chain reaction on the diagnosis of Lung cancer. Methods: 56cases of pathological tissues excised in operation corresponding bronchoalveolar Lavage fluid (BALF) and sputum were obtained from hospitalized patients, among them, 32cases were lung cancer, 24cases were benign Lung diseases. After general dealing the samples, DNA extracting and PCR amp lifiation, the PCR products were dyeing with EB and observed under ultraviolet Lamp. Results: among 32 Lung cancer tissues, 14cases (43.8%) were found abnormal methylation in promoter area of p16 gene, 9cases (64.3%) were obtained methylation in corresponding BALF, 5cases (35.8%) in corresponding sputum, while 24cases of benign Lung diseases were not found methylation neither in tissues excised in operation nor in corresponding BALF and sputum in p16 gene. Conclusion: MSPCR has great speciality on the detection of abnormal methylation in BALF and sputum of patients with lung cancer; MSPCR is a newly and with high potential early diagnostic technique in lung cancer.