title: Characterization of candidate gene copy number alterations in the 11q13 region along with BRAF and NRAS mutations in human melanoma.
abstract: Amplification of the 11q13 chromosomal region is a common event in primary melanomas. Several candidate genes are localized at this sequence; however, their role in melanoma has not been clearly defined. The aim of this study was to develop an accurate method for determining the amplification pattern of six candidate genes that map to this amplicon core and to elucidate the possible relationship between BRAF, NRAS mutations and CCND1 copy number alterations, all of which are key components of the MAP kinase pathway. Characterization of gene copy numbers was performed by quantitative PCR and, as an alternative method, fluorescence in situ hybridization was used to define the CCND1 amplification pattern at the single cell level. Samples with amplified CCND1 (32%) were further analyzed for copy number alterations for the TAOS1, FGF3, FGF19, FGF4 and EMS1 genes. Co-amplification of the CCND1 and TAOS1 was present in 15% of tumors and was more frequent in ulcerated lesions (P=0.017). Furthermore, 56% of primary melanomas had either BRAF or NRAS mutations, but these two mutations were not present in any of the lesions analyzed. Of these cases, 34% also had CCND1 amplification. There was a significant relationship between NRAS activating mutations and UV exposure (P=0.005). We did not find correlations between CCND1 gene amplification status and any of the patients' clinicopathological parameters. However, CCND1 amplification simultaneously with either BRAF or NRAS activation mutations was observed mainly in primary tumors with ulcerated surfaces (P=0.028). We assume that co-amplification of these candidate genes in the 11q13 region or CCND1 gene alterations along with either BRAF or NRAS mutations might be more important for prognosis than the presence of these alterations alone.
authors.affiliation: Department of Preventive Medicine, Faculty of Public Health, Medical and Health Science Center, University of Debrecen, Kassai str. 26, Debrecen, Hungary.
authors.forename: Viktória
authors.initials: V
authors.lastname: Lázár
authors.forename: Szilvia
authors.initials: S
authors.lastname: Ecsedi
authors.forename: Attila G
authors.initials: AG
authors.lastname: Szöllosi
authors.forename: Réka
authors.initials: R
authors.lastname: Tóth
authors.forename: Laura
authors.initials: L
authors.lastname: Vízkeleti
authors.forename: Zsuzsa
authors.initials: Z
authors.lastname: Rákosy
authors.forename: Agnes
authors.initials: A
authors.lastname: Bégány
authors.forename: Róza
authors.initials: R
authors.lastname: Adány
authors.forename: Margit
authors.initials: M
authors.lastname: Balázs
mesh_terms: D000328:Adult; D002880:Chromosomes, Human, Pair 11; D051356:Cortactin / Q000235:genetics; D019938:Cyclin D1 / Q000235:genetics; D005260:Female; D051518:Fibroblast Growth Factor 3 / Q000235:genetics; D051519:Fibroblast Growth Factor 4 / Q000235:genetics; D005346:Fibroblast Growth Factors / Q000235:genetics; D005784:Gene Amplification; D018628:Gene Dosage; D015972:Gene Expression Regulation, Neoplastic; D011905:Genes, ras; D056726:Genetic Association Studies / Q000379:methods; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008545:Melanoma / Q000235:genetics* / Q000556:secondary; D008875:Middle Aged; D009154:Mutation; D009363:Neoplasm Proteins / Q000235:genetics; D009367:Neoplasm Staging; D016133:Polymerase Chain Reaction; D011379:Prognosis; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics*; D015203:Reproducibility of Results; D012878:Skin Neoplasms / Q000235:genetics* / Q000556:secondary; D055815:Young Adult
chemical_list: C530104:CCND1 protein, human; C495868:CTTN protein, human; D051356:Cortactin; C117476:FGF19 protein, human; C496444:FGF3 protein, human; C496445:FGF4 protein, human; D051518:Fibroblast Growth Factor 3; D051519:Fibroblast Growth Factor 4; C465628:LTO1 protein, human; D009363:Neoplasm Proteins; D019938:Cyclin D1; D005346:Fibroblast Growth Factors; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf