Ahrendt S A, Chow J T, Xu L H, Yang S C, Eisenberger C F, Esteller M, Herman J G, Wu L, Decker P A, Jen J, Sidransky D
Department of Surgery, Medical College of Wisconsin, Milwaukee, USA.
J Natl Cancer Inst. 1999 Feb 17;91(4):332-9. doi: 10.1093/jnci/91.4.332.
Conventional cytologic analysis of sputum is an insensitive test for the diagnosis of non-small-cell lung cancer (NSCLC). We have recently demonstrated that polymerase chain reaction (PCR)-based molecular methods are more sensitive than cytologic analysis in diagnosing bladder cancer. In this study, we examined whether molecular assays could identify cancer cells in bronchoalveolar lavage (BAL) fluid.
Tumor-specific oncogene mutations, CpG-island methylation status, and microsatellite alterations in the DNA of cells in BAL fluid from 50 consecutive patients with resectable (stages I through IIIa) NSCLC were assessed by use of four PCR-based techniques.
Of 50 tumors, 28 contained a p53 mutation, and the identical mutation was detected with a plaque hybridization assay in the BAL fluid of 39% (11 of 28) of the corresponding patients. Eight of 19 adenocarcinomas contained a K-ras mutation, and the identical mutation was detected with a mutation ligation assay in the BAL fluid of 50% (four of eight) of the corresponding patients. The p16 gene was methylated in 19 of 50 tumors, and methylated p16 alleles were detected in the BAL fluid of 63% (12 of 19) of the corresponding patients. Microsatellite instability in at least one marker was detected with a panel of 15 markers frequently altered in NSCLC in 23 of 50 tumors; the identical alteration was detected in the BAL fluid of 14% (three of 22) of the corresponding patients. When all four techniques were used, mutations or microsatellite instability was detected in the paired BAL fluid of 23 (53%) of the 43 patients with tumors carrying a genetic alteration.
Although still limited by sensitivity, molecular diagnostic strategies can detect the presence of neoplastic cells in the proximal airway of patients with surgically resectable NSCLC.
痰的传统细胞学分析对非小细胞肺癌（NSCLC）的诊断是一种不敏感的检测方法。我们最近证明，基于聚合酶链反应（PCR）的分子方法在诊断膀胱癌方面比细胞学分析更敏感。在本研究中，我们检测了分子检测方法能否识别支气管肺泡灌洗（BAL）液中的癌细胞。
使用四种基于PCR的技术，对50例连续的可切除（Ⅰ期至Ⅲa期）NSCLC患者的BAL液中细胞的DNA中的肿瘤特异性癌基因突变、CpG岛甲基化状态和微卫星改变进行评估。
50个肿瘤中，28个含有p53突变，在相应患者的39%（28例中的11例）的BAL液中，通过噬斑杂交检测到相同的突变。19例腺癌中有8例含有K-ras突变，在相应患者的50%（8例中的4例）的BAL液中，通过突变连接检测到相同的突变。50个肿瘤中有19个的p16基因发生甲基化，在相应患者的63%（19例中的12例）的BAL液中检测到甲基化的p16等位基因。使用一组在NSCLC中经常改变的15个标记，在50个肿瘤中的23个中检测到至少一个标记的微卫星不稳定性；在相应患者的14%（22例中的3例）的BAL液中检测到相同的改变。当使用所有四种技术时，在43例携带基因改变的肿瘤患者中，有23例（53%）的配对BAL液中检测到突变或微卫星不稳定性。
尽管仍受敏感性的限制，但分子诊断策略可检测出可手术切除的NSCLC患者近端气道中肿瘤细胞的存在。
