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

Hideaki Dote, Kazunori Tsukuda, Shinichi Toyooka, Masaaki Yano, Harvey I. Pass, Nobuyoshi Shimizu

Research output: Contribution to journalArticle

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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.

Original languageEnglish
Pages (from-to)361-363
Number of pages3
JournalOncology Reports
Volume11
Issue number2
Publication statusPublished - Feb 2004

Fingerprint

Codon
Restriction Fragment Length Polymorphisms
Polymerase Chain Reaction
Mutation
Alleles
Malignant Mesothelioma
Protein-Serine-Threonine Kinases
Point Mutation
Ovarian Neoplasms
Exons
Melanoma
Thyroid Gland
Cell Line
Growth
Neoplasms

Keywords

  • BRAF
  • Enriched PCR-RFLP
  • Malignant pleural mesothelioma

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Dote, H., Tsukuda, K., Toyooka, S., Yano, M., Pass, H. I., & Shimizu, N. (2004). Mutation analysis of the BRAF codon 599 in malignant pleural mesothelioma by enriched PCR-RFLP. Oncology Reports, 11(2), 361-363.

Mutation analysis of the BRAF codon 599 in malignant pleural mesothelioma by enriched PCR-RFLP. / Dote, Hideaki; Tsukuda, Kazunori; Toyooka, Shinichi; Yano, Masaaki; Pass, Harvey I.; Shimizu, Nobuyoshi.

In: Oncology Reports, Vol. 11, No. 2, 02.2004, p. 361-363.

Research output: Contribution to journalArticle

Dote, H, Tsukuda, K, Toyooka, S, Yano, M, Pass, HI & Shimizu, N 2004, 'Mutation analysis of the BRAF codon 599 in malignant pleural mesothelioma by enriched PCR-RFLP', Oncology Reports, vol. 11, no. 2, pp. 361-363.
Dote, Hideaki ; Tsukuda, Kazunori ; Toyooka, Shinichi ; Yano, Masaaki ; Pass, Harvey I. ; Shimizu, Nobuyoshi. / Mutation analysis of the BRAF codon 599 in malignant pleural mesothelioma by enriched PCR-RFLP. In: Oncology Reports. 2004 ; Vol. 11, No. 2. pp. 361-363.
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