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 ( MSPCR)在肺癌临床诊断中的应用价值。方法 :选取 5 6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 ,2 4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 ,1 4例 ( 4 3.8% )在 p1 6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中检出甲基化存在 ,5例 ( 35 .8% )在相应的痰标本中也检出甲基化存在。良性肺部疾病 2 4例 ,其中肺囊肿 1 0例 ,肺结核 1 4例 ,无论在手术切除标本还是 BALF和痰标本中均未检出 p1 6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 ,是一项很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:56 cases of pathological tissues excised in operation, corresponding bronchoalveolar lavage fluid(BALF) and sputum were obtained from hospitalized patients, among whom,32 cases were lung cancer,24 cases were benign Lung diseases.After DNA extracting and PCR amplifiation,the PCR products were dyed with EB and observed under ultraviolet lamp. Results:Among 32 lung cancer tissues,14 cases(43.8%) showed abnormal methylation in promoter area of p16 gene,9 cases(64.3%) were obtained methylation in corresponding BALF,5 cases(35.8%) in corresponding sputum,while 24 cases of benign lung diseases did not show methylation either in tissues excised in operation or in corresponding BALF and sputum. Conclusion:MSPCR is specific in the detection of abnormal methylation in BALF and sputum of patients with lung cancer; MSPCR is a new and highly potential early diagnostic technique in lung cancer.