Detection of codon 61 point mutations of the K-ras gene in lung and colorectal cancers by enriched PCR

Oncol Rep. Sep-Oct 2003;10(5):1455-9. doi: 10.3892/or.10.5.1455.

Abstract

Mutation of the Kirsten ras (K-ras) gene is one of most common alterations in solid tumors including lung and colorectal cancers. We developed new enriched PCR-RFLP assay to detect mutations of K-ras codon 61 at the 1st and 2nd letters and non-enriched PCR-RFLP assay to detect the 3rd letter mutation. One mutant allele among 10(3) wild-type alleles was detected by enriched PCR-RFLP assay, while one mutant in 10 wild-type alleles was detected by non-enriched PCR-RFLP assay for codon 61 3rd letter. We then examined K-ras codon 12, 13 and 61 mutations in lung and colorectal cancers using these assays. K-ras codon 12 mutation was detected in 10 of 109 (9%) lung cancer and 19 of 83 (23%) colorectal cancer cases. K-ras codon 13 mutation was detected in 2 of 83 (2%) colorectal and 0 of 109 NSCLC cases, respectively. There was no K-ras codon 61 mutation in either type of cancer. Our results demonstrate that enriched PCR-RFLP is a sensitive assay to detect K-ras codon 61 mutation, however, it was extremely rare in lung and colorectal cancers, suggesting organ-specific pathways in mutagenesis of the ras gene family.

MeSH terms

  • Aged
  • Alleles
  • Codon*
  • Colorectal Neoplasms / genetics*
  • DNA
  • Exons
  • Female
  • Genes, ras*
  • Humans
  • Lung Neoplasms / genetics*
  • Male
  • Middle Aged
  • Mutation*
  • Point Mutation*
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Restriction Fragment Length

Substances

  • Codon
  • DNA