Kimura S, Fukuchi K, Takeda S, Shin S, Saito D, Akamatsu S, Himeji Y, Chen G L, Watanabe H, Takagi Y
Department of Clinical Pathology, Showa University School of Medicine, Tokyo.
Rinsho Byori. 1995 Jul;43(7):718-23.
Sputum cells from 44 lung cancer patients (22 adenocarcinoma, 19 squamous cell carcinoma and 3 small cell carcinoma cases) were examined to contain mutant K-ras genes. The mutant-allele-specific amplification (MASA) method was used for detection of K-ras point mutations. The reaction was designed to amplify only mutant codons with polymerase chain reaction (PCR). Codons 12, 13 and 61 were studied with synthesized primers. Two cases (primary adenocarcinoma of the lung and pancreas adenocarcinoma with lung metastasis) were positive for point mutations at codon 12. The mutation of GGT to GTT was found in the first case and to GAT in the second one. Further experiments with colony hybridization revealed the mutations were found in 1% of alleles in both cases. There were no point mutations found at codon 13 nor 61. Our experiments showed it is possible to detect K-ras mutation in sputum cells from lung cancer patients, with MASA method.
对44例肺癌患者（22例腺癌、19例鳞状细胞癌和3例小细胞癌）的痰液细胞进行检测，以确定是否含有突变的K-ras基因。采用突变等位基因特异性扩增（MASA）方法检测K-ras点突变。该反应设计为仅用聚合酶链反应（PCR）扩增突变密码子。用合成引物研究密码子12、13和61。2例（原发性肺腺癌和肺转移的胰腺腺癌）密码子12存在点突变阳性。第一例中发现GGT突变为GTT，第二例中突变为GAT。进一步的菌落杂交实验表明，两例中均有1%的等位基因存在突变。在密码子13和61处未发现点突变。我们的实验表明，采用MASA方法能够检测肺癌患者痰液细胞中的K-ras突变。
