title: High frequency of homozygosity of the HLA region in melanoma cell lines reveals a pattern compatible with extensive loss of heterozygosity.
abstract: Malignant transformation of cells is frequently associated with abnormalities in human leukocyte antigen (HLA) expression. MHC class I loss or down-regulation in cancer cells is a major immune escape route used by a large variety of human tumours to evade antitumour immune responses mediated by cytotoxic T lymphocytes. The goal of our study was to explore HLA genotyping and phenotyping in a variety of melanoma tumour cell lines. A total of 91 melanoma cell lines were characterised for HLA class I and II genotype. In addition, 61 out of the 91 cell lines were also analysed for HLA class I and II cell surface molecule expression by flow cytometry. Unexpectedly, we found that 19.7% of the melanoma cell lines were homozygous for HLA class I genotypes, sometimes associated with HLA class II homozygosity (8.79%) and sometimes not (10.98%). The frequency of homozygosity was significantly higher compared with the control groups (1.6%). To identify the reasons underlying the high frequency of HLA homozygosity we searched for genomic deletions using eight pairs of highly polymorphic microsatellite markers covering the entire extended HLA complex on the short arm of chromosome 6. Our results were compatible with hemizygous deletions and suggest that loss of heterozygosity on chromosome arm 6p is a common feature in melanoma cell lines. In fact, although autologous normal DNA from the patients was not available and could not be tested, the retention in some cases of heterozygosity for a number of microsatellite markers would indicate a hemizygous deletion. In the rest of the cases, markers at 6p and 6q showed a single allele pattern indicating the probable loss of part or the whole of chromosome 6. These results led us to conclude that loss of heterozygosity in chromosome 6 is nonrandom and is possibly an immunologically relevant event in human malignant melanoma. Other well-established altered HLA class I phenotypes were also detected by flow cytometry that correspond to HLA class I total loss and HLA-ABC and/or specific HLA-B locus down-regulation.
authors.affiliation: Departamento de Análisis Clínicos, Hospital Universitario Virgen de las Nieves, Universidad de Granada, Avd. Fuerzas Armadas 2, 18014 Granada, Spain.
authors.forename: Teresa
authors.initials: T
authors.lastname: Rodriguez
authors.forename: Rosa
authors.initials: R
authors.lastname: Méndez
authors.forename: Chrissy H
authors.initials: CH
authors.lastname: Roberts
authors.forename: Francisco
authors.initials: F
authors.lastname: Ruiz-Cabello
authors.forename: I Anthony
authors.initials: IA
authors.lastname: Dodi
authors.forename: Miguel Angel
authors.initials: MA
authors.lastname: López Nevot
authors.forename: Laura
authors.initials: L
authors.lastname: Paco
authors.forename: Isabel
authors.initials: I
authors.lastname: Maleno
authors.forename: Steven G E
authors.initials: SG
authors.lastname: Marsh
authors.forename: Graham
authors.initials: G
authors.lastname: Pawelec
authors.forename: Federico
authors.initials: F
authors.lastname: Garrido
mesh_terms: D002896:Chromosomes, Human, Pair 6 / Q000235:genetics*; D015536:Down-Regulation; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D005805:Genes, MHC Class I / Q000235:genetics*; D005802:Genes, MHC Class II / Q000235:genetics*; D005838:Genotype; D006720:Homozygote; D006801:Humans; D019656:Loss of Heterozygosity; D008545:Melanoma / Q000235:genetics* / Q000276:immunology; D018895:Microsatellite Repeats; D010641:Phenotype; D012878:Skin Neoplasms / Q000235:genetics / Q000276:immunology; D014407:Tumor Cells, Cultured