title: Chromosome 17 allelic loss and NF1-GRD mutations do not play a significant role as molecular mechanisms leading to melanoma tumorigenesis.
abstract: Allelic loss in human cutaneous melanoma has been detected on chromosomes 1p, 6q, 9p, 10q, and 11q. Chromosome 17 contains important tumor suppressor genes such as p53, NM23, and neurofibromatosis type 1 (NF1), which have been implicated in melanoma tumorigenesis. The role of p53 has already been studied by a number of laboratories, showing contrasting results. In the present study, two restriction fragment length polymorphism (RFLP) probes for the NM23 and NF1 genes, together with five other RFLP and four variable number of tandem repeat chromosome 17 probes, were investigated at the loss of heterozygosity (LOH) level in a Southern blot-based assay. The NF1 gene was also tested for LOH by a polymerase chain reaction (PCR)-based approach in two different experiments, using a dinucleotide repeat polymorphic probe at locus D17S250 (17q11.2-q12), and an Alu probe intragenic to the NF1 gene (17q11.2). A PCR single-strand conformation polymorphism assay was included in the study for mutation detection at the NF1-GTPase-activating protein-related domain (GRD). A total of 68 melanocytic tumors were analyzed. LOH was detected in 9 of 87 informative cases (10%). LEW301 (17p11.2-pcen) presented the highest LOH frequency (22%). NM23 showed LOH in 17% of the informative cases, while NF1 did not show either LOH in the Southern blot- and PCR-based experiments or mutations at the NF1-GRD. These results are in concordance with those of previous smaller studies, but when compared with higher LOH frequencies obtained from other chromosomes, these findings indicate that the LOH values found in our study can most likely be attributed to background effect. Thus, chromosome 17 LOH is likely to play and unimportant role as a genetic event in melanoma tumorigenesis. Nevertheless, NF1 merits further study, since homozygous deletions have been detected at this locus in melanoma cell lines.
authors.affiliation: Instituto de Investigaciones Biomedicas, C.S.I.C., Arturo Duperier 4, Madrid, Spain.
authors.forename: L
authors.initials: L
authors.lastname: Gómez
authors.forename: M P
authors.initials: MP
authors.lastname: Rubio
authors.forename: M T
authors.initials: MT
authors.lastname: Martin
authors.forename: J J
authors.initials: JJ
authors.lastname: Vázquez
authors.forename: M
authors.initials: M
authors.lastname: Idoate
authors.forename: G
authors.initials: G
authors.lastname: Pastorfide
authors.forename: A
authors.initials: A
authors.lastname: Pestaña
authors.forename: B R
authors.initials: BR
authors.lastname: Seizinger
authors.forename: R L
authors.initials: RL
authors.lastname: Barnhill
authors.forename: J S
authors.initials: JS
authors.lastname: Castresana
mesh_terms: D000483:Alleles; D002886:Chromosomes, Human, Pair 17 / Q000235:genetics*; D004273:DNA, Neoplasm / Q000235:genetics; D020690:GTPase-Activating Proteins; D016514:Genes, Neurofibromatosis 1; D006579:Heterozygote; D006801:Humans; D008545:Melanoma / Q000209:etiology / Q000235:genetics*; D009154:Mutation; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D011506:Proteins / Q000235:genetics*
chemical_list: D004273:DNA, Neoplasm; D020690:GTPase-Activating Proteins; D011506:Proteins