title: Molecular characterization of human cutaneous melanoma-derived cell lines.
abstract: BACKGROUND
  Several studies have demonstrated that different genetic profiles contribute to melanoma development and progression.
  
  
  METHODS
  To evaluate the existence of different molecular aberration patterns in melanoma associated with v-raf murine sarcoma viral oncogene homolog B1 (BRAF) or 9p21 locus alterations, eleven patient-derived melanoma cell lines were characterized. Multiplex ligation probe amplification (MLPA) was used to detect chromosomal alterations. Single- strand conformation analysis and sequencing were performed to study BRAF, neuroblastoma RAS viral (v-ras) oncogene homolog (NRAS), v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog (c-KIT), melanocortin 1 receptor (alpha melanocyte stimulating hormone receptor) (MC1R), cyclin-dependent kinase inhibitor 2A (CDKN2A) and cyclin-dependent kinase 4 (CDK4) genes.
  
  
  RESULTS
  BRAFV600E mutation was detected in 54% of cell lines. NRAS was mutated in one cell line also carrying multiple copies of NRAS. All cell lines with MC1R variants harboured BRAFV600E. Concurrent loss of MUTYH (1p33), gains of c-MYC (8q24) and of CDK6 (7q21) were found to be significantly associated in cell lines (45%) that harboured biallelic 9p21 deletions including CDKN2B-CDKN2A-MTAP.
  
  
  CONCLUSIONS
  These data suggest the existence of a specific pattern of somatic alterations in genes that are involved in DNA repair (MUTYH) and in cell cycle regulation (c-MYC, CDK6, CDKN2A and CDKN2B). Interestingly, all MC1R variants were associated with BRAFV600E and all cell lines from visceral metastases harboured BRAFV600E.
authors.affiliation: Melanoma Unit of the Department of Dermatology, Hospital Clínic of Barcelona, IDIBAPS, Barcelona University, Barcelona, Spain. zigheog@gmail.com
authors.forename: Zighereda
authors.initials: Z
authors.lastname: Ogbah
authors.forename: Joan Anton
authors.initials: JA
authors.lastname: Puig-Butille
authors.forename: Federico
authors.initials: F
authors.lastname: Simonetta
authors.forename: Celia
authors.initials: C
authors.lastname: Badenas
authors.forename: Remedios
authors.initials: R
authors.lastname: Cervera
authors.forename: Jordi
authors.initials: J
authors.lastname: Milá
authors.forename: Daniel
authors.initials: D
authors.lastname: Benitez
authors.forename: Josep
authors.initials: J
authors.lastname: Malvehy
authors.forename: Ramon
authors.initials: R
authors.lastname: Vilella
authors.forename: Susana
authors.initials: S
authors.lastname: Puig
mesh_terms: D001483:Base Sequence; D045744:Cell Line, Tumor; D051358:Cyclin-Dependent Kinase 4 / Q000235:genetics; D017931:DNA Primers; D011905:Genes, ras; D006801:Humans; D008545:Melanoma / Q000235:genetics* / Q000473:pathology; D009154:Mutation; D016133:Polymerase Chain Reaction; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics; D044102:Receptor, Melanocortin, Type 1 / Q000235:genetics; D012878:Skin Neoplasms / Q000235:genetics* / Q000473:pathology
chemical_list: D017931:DNA Primers; D044102:Receptor, Melanocortin, Type 1; D019009:Proto-Oncogene Proteins c-kit; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf; C495900:CDK4 protein, human; D051358:Cyclin-Dependent Kinase 4