title: Analysis of c-KIT exon 11 mutations in canine gastrointestinal stromal tumours.
abstract: The aim of this study was to determine the type and frequency of c-KIT exon 11 mutations in canine gastrointestinal stromal tumours (GISTs) and investigate the association between the c-KIT mutation status and KIT immunohistochemical staining pattern. Mutations in exon 11 of c-KIT were examined in 46 formalin-fixed paraffin-embedded canine GISTs using PCR of genomic DNA and reverse transcription-PCR (RT-PCR) of cDNA. Exon 11 c-KIT mutations were detected in 15/46 (32.6%) cases by conventional PCR and 34/46 (73.9%) cases by RT-PCR; the mutation detection rate was significantly higher for RT-PCR (P = 0.004, Fisher's exact test). Ten different mutations, including deletion, internal tandem duplication and point mutations, were identified by RT-PCR. Immunohistochemistry was performed using an anti-KIT antibody; diffuse KIT staining was detected in the tumour cell cytoplasm in 32/46 (69.6%) cases and partial or stippled cytoplasmic staining of KIT was observed in 14/46 (30.4%) cases. Neither pattern was significantly associated with c-KIT exon 11 mutation status (P = 1.000, chi-square test). These data indicate that c-KIT exon 11 mutations occur frequently in canine GISTs, similar to human GISTs; however, there is no association between c-KIT mutations and the KIT expression pattern in canine GISTs. This study suggests that RT-PCR is more sensitive than conventional PCR for the detection of c-KIT mutations in canine GISTs.
authors.affiliation: Nasunogahara Animal Clinic, 2-3574-98, Asaka, Ohtawara, Tochigi 324-0043, Japan. Electronic address: volkerhall@me.com.
authors.forename: M
authors.initials: M
authors.lastname: Takanosu
authors.affiliation: North Lab, 2-8-35, Hondori, Shiroisi-ku, Sapporo, Hokkaido 003-0027, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Amano
authors.affiliation: North Lab, 2-8-35, Hondori, Shiroisi-ku, Sapporo, Hokkaido 003-0027, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Kagawa
mesh_terms: D000818:Animals; D004283:Dog Diseases / Q000235:genetics*; D004285:Dogs; D005091:Exons; D046152:Gastrointestinal Stromal Tumors / Q000235:genetics / Q000662:veterinary*; D006801:Humans; D007150:Immunohistochemistry / Q000662:veterinary; D009154:Mutation; D059645:Mutation Rate; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D013045:Species Specificity
chemical_list: D019009:Proto-Oncogene Proteins c-kit
keywords: Canine; Gastrointestinal stromal tumours; Immunohistochemistry; Mutation; c-KIT