title: Comparative analysis of BRAF, NRAS and c-KIT mutation status between tumor tissues and autologous tumor cell-lines of stage III/IV melanoma.
abstract: In the last decade, advances in molecular biology have provided evidence of the genotypic heterogeneity of melanoma. We analysed BRAF, NRAS and c-KIT alterations in tissue samples from 63 stage III/IV melanoma patients and autologous cell-lines, using either allele-specific or quantitative PCR. The expression of BRAF V600E protein was also investigated using an anti-BRAF antibody in the same tissue samples. 81% of FFPE samples and tumor cell-lines harboured a genetic alteration in either BRAF (54%) or NRAS (27%) oncogenes. There was a strong concordance (100%) between tissue samples and tumor cell-lines. The BRAF V600E mutant-specific antibody showed high sensitivity (96%) and specificity (100%) for detecting the presence of a BRAF V600E mutation. The correlation was of 98% between PCR and immunohistochemistry results for BRAF mutation. These results suggest that BRAF and NRAS mutation status of tumor cells is not affected by culture conditions.
authors.affiliation: CRCNA, INSERM U892, CNRS 6299, Nantes Cedex 01, France.
authors.forename: Anne-Chantal
authors.initials: AC
authors.lastname: Knol
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Pandolfino
authors.forename: Audrey
authors.initials: A
authors.lastname: Vallée
authors.forename: Frédérique
authors.initials: F
authors.lastname: Nguyen
authors.forename: Virginie
authors.initials: V
authors.lastname: Lella
authors.forename: Amir
authors.initials: A
authors.lastname: Khammari
authors.forename: Marc
authors.initials: M
authors.lastname: Denis
authors.forename: Anne-Laure
authors.initials: AL
authors.lastname: Puaux
authors.forename: Brigitte
authors.initials: B
authors.lastname: Dréno
mesh_terms: D000328:Adult; D000368:Aged; D000483:Alleles; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D004252:DNA Mutational Analysis; D005260:Female; D020558:GTP Phosphohydrolases / Q000235:genetics* / Q000378:metabolism; D005838:Genotype; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008545:Melanoma / Q000378:metabolism* / Q000473:pathology; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D008875:Middle Aged; D009154:Mutation; D009857:Oncogenes; D016133:Polymerase Chain Reaction; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics* / Q000378:metabolism; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics* / Q000378:metabolism; D012680:Sensitivity and Specificity; D012878:Skin Neoplasms / Q000378:metabolism* / Q000473:pathology; D055815:Young Adult
chemical_list: D008565:Membrane Proteins; D019009:Proto-Oncogene Proteins c-kit; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf; D020558:GTP Phosphohydrolases; C579846:NRAS protein, human
keywords: genetic alterations; metastatic melanoma; tumor cell-lines