[Detection of K-ras mutation from malignant cells in sputum with mutant-allele specific amplification (MASA) method]

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.