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Anticancer Res. 2016 Apr;36(4):1571-9.

Rapid and Specific Screening Assay for KRAS Oncogene Mutation by a Novel Gene Amplification Method.

Author information

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.

KEYWORDS:

KRAS mutation; LAMP; molecular targeted therapy; stratification biomarker

PMID:
27069133
[Indexed for MEDLINE]

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