title: Rapid detection of mutations in the BRAF gene using real-time polymerase chain reaction and melting curve analysis.
abstract: The BRAF gene is mutated in 66% of melanomas and less frequently in various human cancers. More than 80% of these mutations are T to A transversions at nucleotide 1796 (T1796A), leading to a substitution of glutamic acid for valine at amino acid 599 (V599E). We established a new method for rapidly detecting V599E mutations using real-time polymerase chain reaction and melting curve analysis. Furthermore, we examined mutations in gastrointestinal cancer cell lines using this method. We found a mutation in 1 of 12 (8%) colorectal cancer cell lines, but no mutation was detected in 9 gastric cancer cell lines. These results suggest that the BRAF mutation is unlikely to be involved in gastric carcinogenesis.
authors.affiliation: Division of Gastroenterology, The Institute for Adult Diseases, Asahi Life Foundation, 1-9-14, Nishi-Shinjuku, Shinjuku-ku, Tokyo 160-0023, Japan. t-ikenoue@asahi-life.or.jp
authors.forename: Tsuneo
authors.initials: T
authors.lastname: Ikenoue
authors.forename: Yohko
authors.initials: Y
authors.lastname: Hikiba
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Kanai
authors.forename: Hideaki
authors.initials: H
authors.lastname: Ijichi
authors.forename: Goichi
authors.initials: G
authors.lastname: Togo
authors.forename: Miki
authors.initials: M
authors.lastname: Ohta
authors.forename: Hirotsugu
authors.initials: H
authors.lastname: Watabe
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Yamaji
authors.forename: Makoto
authors.initials: M
authors.lastname: Okamoto
authors.forename: Jun
authors.initials: J
authors.lastname: Aragaki
authors.forename: Masayuki
authors.initials: M
authors.lastname: Matsumura
authors.forename: Takao
authors.initials: T
authors.lastname: Kawabe
authors.forename: Masao
authors.initials: M
authors.lastname: Omata
mesh_terms: D019943:Amino Acid Substitution; D015179:Colorectal Neoplasms / Q000235:genetics*; D017931:DNA Primers / Q000737:chemistry; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification*; D005770:Gastrointestinal Neoplasms / Q000235:genetics*; D006801:Humans; D009154:Mutation / Q000235:genetics*; D009691:Nucleic Acid Denaturation; D048493:Proto-Oncogene Proteins B-raf; D019908:Proto-Oncogene Proteins c-raf / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014407:Tumor Cells, Cultured
chemical_list: D017931:DNA Primers; D004273:DNA, Neoplasm; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf; D019908:Proto-Oncogene Proteins c-raf