title: Peptide nucleic acid-locked nucleic acid polymerase chain reaction clamp-based detection test for gefitinib-refractory T790M epidermal growth factor receptor mutation.
abstract: Mutations in the epidermal growth factor receptor (EGFR) are observed in a fraction of non-small-cell lung cancers (NSCLS). EGFR mutation-positive NSCLS responds to gefitinib. Secondary T790M mutation confers gefitinib resistance to NSCLS. A detection test for the T790M mutation was designed based on the peptide nucleic acid-locked nucleic acid polymerase chain reaction clamp method. The specificity and sensitivity of the test were both greater than 0.99. The test revealed that only a small population of the PC-13 cells carried the T790M mutation. The test also revealed that the T790M mutation was found in none of 151 NSCLC specimens obtained before gefitinib treatment, whereas it was found in four of four specimens obtained from NSCLS that had become refractory to gefitinib. In one patient in whom the L858R-positive EGFR allele was amplified to multiple copies, an L858R-T790M double-mutant allele emerged during the gefitinib therapy. This allele was expressed highly. The T790M mutation detection test based on the peptide nucleic acid-locked nucleic acid polymerase chain reaction clamp method is sensitive and specific, and is applicable to clinical practice. It detects T790M-positive cells in the course of gefitinib treatment, and thus will help to devise therapies effective for T790M-positive NSCLS.
authors.affiliation: Department of Respiratory Medicine, Saitama Medical University, 38 Morohongo, Moroyama-machi, Saitama 350-0495, Japan.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Miyazawa
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Tanaka
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Nagai
authors.forename: Masaru
authors.initials: M
authors.lastname: Matsuoka
authors.forename: Akihisa
authors.initials: A
authors.lastname: Sutani
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Udagawa
authors.forename: Jialing
authors.initials: J
authors.lastname: Zhang
authors.forename: Takashi
authors.initials: T
authors.lastname: Hirama
authors.forename: Yoshitake
authors.initials: Y
authors.lastname: Murayama
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Koyama
authors.forename: Kenji
authors.initials: K
authors.lastname: Ikebuchi
authors.forename: Makoto
authors.initials: M
authors.lastname: Nagata
authors.forename: Minoru
authors.initials: M
authors.lastname: Kanazawa
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Nukiwa
authors.forename: Seiichi
authors.initials: S
authors.lastname: Takenoshita
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Kobayashi
authors.forename: Koichi
authors.initials: K
authors.lastname: Hagiwara
mesh_terms: D000970:Antineoplastic Agents / Q000627:therapeutic use*; D001483:Base Sequence; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy* / Q000235:genetics; D045744:Cell Line, Tumor; D004252:DNA Mutational Analysis / Q000379:methods*; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D066246:ErbB Receptors / Q000235:genetics*; D000077156:Gefitinib; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000235:genetics*; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry; D020135:Peptide Nucleic Acids / Q000737:chemistry; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011799:Quinazolines / Q000627:therapeutic use*; D012680:Sensitivity and Specificity
chemical_list: D000970:Antineoplastic Agents; D009841:Oligonucleotides; D020135:Peptide Nucleic Acids; D011799:Quinazolines; C477371:locked nucleic acid; D066246:ErbB Receptors; D000077156:Gefitinib