Rapid detection of mutations in the BRAF gene using real-time polymerase chain reaction and melting curve analysis.
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.
Ikenoue
Tsuneo
T
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
Hikiba
Yohko
Y
Kanai
Fumihiko
F
Ijichi
Hideaki
H
Togo
Goichi
G
Ohta
Miki
M
Watabe
Hirotsugu
H
Yamaji
Yutaka
Y
Okamoto
Makoto
M
Aragaki
Jun
J
Matsumura
Masayuki
M
Kawabe
Takao
T
Omata
Masao
M
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
D017931:DNA Primers; D004273:DNA, Neoplasm; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf; D019908:Proto-Oncogene Proteins c-raf