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Mutation analysis of the BRAF codon 599 in malignant pleural mesothelioma by enriched PCR-RFLP.

Abstract
BRAF encodes a RAS-regulated serine/threonine kinase that mediates the pathway for cell growth and malignant transformation. Point mutations of BRAF were reported recently in 66% of melanomas, over 30% of thyroid papillary and low-grade ovarian cancers, and a smaller percentage of other human cancers. Mutations in malignant cells were reported to occur only in exons 11 and 15. Among these mutations, BRAF V599E is most frequent and proved to invert its transcript to the dominant active form. To exclude the interference of co-existing normal cells in clinical samples, we developed a new enriched PCR-RFLP assay for detecting mutations of BRAF codon 599 mutation. The sensitivity of this assay was examined to find that one mutant allele among 10(2) wild-type alleles could be detected. We applied this method for 53 cases of primary malignant pleural mesotheliomas (MPMs) and 6 cell lines and found no mutations in these samples. Our results demonstrate that the developed enriched PCR-RFLP is a sensitive assay to detect BRAF codon 599 mutation. However, it may be a rare type of mutation in MPMs. Our new assay is useful and can be applied for screening of BRAF codon 599 mutation in various kinds of clinical samples.
AuthorsHideaki Dote, Kazunori Tsukuda, Shinichi Toyooka, Masaaki Yano, Harvey I Pass, Nobuyoshi Shimizu
JournalOncology reports (Oncol Rep) Vol. 11 Issue 2 Pg. 361-3 (Feb 2004) ISSN: 1021-335X [Print] Greece
PMID14719068 (Publication Type: Journal Article)
Chemical References
  • Codon
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf
  • Proto-Oncogene Proteins c-raf
Topics
  • Cell Division
  • Codon (genetics)
  • DNA Mutational Analysis
  • Humans
  • Mesothelioma (genetics, pathology, surgery)
  • Pleural Neoplasms (genetics, pathology, surgery)
  • Point Mutation (genetics)
  • Polymerase Chain Reaction (methods)
  • Polymorphism, Restriction Fragment Length
  • Proto-Oncogene Proteins B-raf
  • Proto-Oncogene Proteins c-raf (genetics)
  • Sensitivity and Specificity

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