A modified mutagenic PCR-RFLP method for K-ras codon 12 and 13 mutations detection in NSCLC patients

Citation
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
Citations number
21
Categorie Soggetti
Molecular Biology & Genetics
Journal title
MOLECULAR AND CELLULAR PROBES
ISSN journal
08908508 → ACNP
Volume
15
Issue
5
Year of publication
2001
Pages
243 - 247
Database
ISI
SICI code
0890-8508(200110)15:5<243:AMMPMF>2.0.ZU;2-F
Abstract
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.