A. Hatzaki et al., A modified mutagenic PCR-RFLP method for K-ras codon 12 and 13 mutations detection in NSCLC patients, MOL CELL PR, 15(5), 2001, pp. 243-247
Evidence from many investigators has shown that mutations in the first exon
of K-ras gene occur at elevated frequencies in lung, pancreatic and colon
carcinoma and seem to be of prognostic importance. The aim of this study wa
s to develop an effective method for the detection of K-ras mutations in co
dons 12 and 13 in non-small-cell lung cancer (NSCLC) patients in order to i
nvestigate correlation with clinical outcome.
DNA was extracted from tumour and neighbouring non-neoplastic lung tissues
from 70 patients and screened for codon 12 and 13 mutations. We applied a m
utagenic PCR-restriction fragment length polymorphism for both codon 12 and
13 mutation detection. Codon 12 mutation was identified in 20% of NSCLC pa
tients, whereas no codon 13 mutation was detected. As expected, the respect
ive non-neoplastic tissues exhibited no mutations. We observed an increased
codon 12 mutation prevalence in adenocarcinoma comparing to other types of
carcinomas. Follow-up for 29 patients with a mean time of 12 months indica
tes an increased relapse rate in NSCLC patients with the K-ras codon 12 mut
ation. Furthermore, a trend towards increased percentage of mutant samples
was observed in the advanced stage group of patients.
We provide evidence that our approach is a fast and reliable method for scr
eening K-ras exon 1 mutations in tumour samples from NSCLC patients. (C) 20
01 Academic Press.