Akira Kumasaka1, Naoyuki Matsumoto2, Shotaro Mukae3, Taiichi Kitano3, Hiroyasu Noguchi4, Hidero Ohki4, Kazuo Komiyama3, Tomohiro Ando1. 1. Department of Oral and Maxillofacial Surgery, Tokyo Women's Medical University, School of Medicine, Shinjyuku-ku, Tokyo, Japan. 2. Department of Pathology, Nihon University School of Dentistry, Chiyoda-ku, Tokyo, Japan matsumoto.naoyuki@nihon-u.ac.jp. 3. Department of Pathology, Nihon University School of Dentistry, Chiyoda-ku, Tokyo, Japan. 4. Department of Oral and Maxillofacial Surgery, Nihon University School of Dentistry, Chiyoda-ku, Tokyo, Japan.
Abstract
BACKGROUND: Epidermal growth factor receptor (EGFR) is a target of molecular therapeutics for colorectal cancer. However, mutation of Kirsten rat sarcoma viral oncogene homolog (KRAS) gene at codons 12 and 13 attenuates the therapeutic effect of anti-EGFR therapies. Therefore, the detection of KRAS gene mutation is important for therapeutic decision-making. MATERIALS AND METHODS: KRAS gene mutation at codons 12 (c.34G>T, c.35G>C, c.35G>A) and 13 (c.38G>A) in six cancer cell lines were investigated. A loop-mediated isothermal amplification-based procedure was developed that employed peptide nucleic acid to suppress amplification of the wild-type allele. RESULTS: This mutation-oriented gene-amplification procedure can amplify the DNA fragment of the KRAS gene with codon 12 and codon 13 mutation within 30 min. Moreover, boiled cells can work as template resources. CONCLUSION: This newly developed procedure can be useful for patient stratification for anti-EGFR therapies. Copyright
BACKGROUND:Epidermal growth factor receptor (EGFR) is a target of molecular therapeutics for colorectal cancer. However, mutation of Kirsten rat sarcoma viral oncogene homolog (KRAS) gene at codons 12 and 13 attenuates the therapeutic effect of anti-EGFR therapies. Therefore, the detection of KRAS gene mutation is important for therapeutic decision-making. MATERIALS AND METHODS:KRAS gene mutation at codons 12 (c.34G>T, c.35G>C, c.35G>A) and 13 (c.38G>A) in six cancer cell lines were investigated. A loop-mediated isothermal amplification-based procedure was developed that employed peptide nucleic acid to suppress amplification of the wild-type allele. RESULTS: This mutation-oriented gene-amplification procedure can amplify the DNA fragment of the KRAS gene with codon 12 and codon 13 mutation within 30 min. Moreover, boiled cells can work as template resources. CONCLUSION: This newly developed procedure can be useful for patient stratification for anti-EGFR therapies. Copyright