肺癌组织、支气管肺泡灌洗液及痰标本中p16基因甲基化特异性PCR检测

Detection of aberrant methylation of p16 gene in lung tumor tissues,corresponding bronchoalveolar lavage fluid, and sputum by methylation-specific PCR

目的 :探讨肺癌组织、支气管灌洗液及痰标本中p16基因甲基化特异性PCR检测的临床应用价值。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 ,14例 (4 3.8% )在p16基因启动子区域呈现异常甲基化 ,其中 9例 (6 4 .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。 2 4例良性肺部疾病 ,其中肺囊肿 10例 ,肺结核 14例 ,无论在手术切除标本还是BALF和痰标本中均未检出p16基因甲基化存在。结论 :MSP技术对肺癌患者BALF及痰标本中P16基因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。

Aim:To explore the clinical value of methylation specific PCR (MSP) on the diagnosis of Lung cancer. Methods:Pathological tissues taken in operation, corresponding bronchoalveolar lavage fluid (BALF), and sputum were obtained from 56 hospitalized patients, among whom 32 were lung cancer, and 24 were benign lung diseases.After DNA extracting and PCR amplification, the PCR products were stained with EB and observed under ultraviolet lamp.Results:Among 32 lung cancer tissues,14(43.8%) were found abnormal methylation in promoter area of p16 gene,9(64.3%), in corresponding BALF, and 5(35.8%), in corresponding sputum. While among 24 cases of benign lung diseases no p16 gene methylation was found in pathological tissues taken in operation, corresponding BALF or sputum. Conclusion:MSP has specificity on the detection of abnormal methylation of P16 gene in BALF and sputum of patients with lung cancer. This suggests that MSP might be a promising technique for lung cancer early diagnosis