title: Combined M-FISH and CGH analysis allows comprehensive description of genetic alterations in neuroblastoma cell lines.
abstract: Cancer cell lines are essential gene discovery tools and have often served as models in genetic and functional studies of particular tumor types. One of the future challenges is comparison and interpretation of gene expression data with the available knowledge on the genomic abnormalities in these cell lines. In this context, accurate description of these genomic abnormalities is required. Here, we show that a combination of M-FISH with banding analysis, standard FISH, and CGH allowed a detailed description of the genetic alterations in 16 neuroblastoma cell lines. In total, 14 cryptic chromosome rearrangements were detected, including a balanced t(2;4)(p24.3;q34.3) translocation in cell line NBL-S, with the 2p24 breakpoint located at about 40 kb from MYCN. The chromosomal origin of 22 marker chromosomes and 41 cytogenetically undefined translocated segments was determined. Chromosome arm 2 short arm translocations were observed in six cell lines (38%) with and five (31%) without MYCN amplification, leading to partial chromosome arm 2p gain in all but one cell line and loss of material in the various partner chromosomes, including 1p and 11q. These 2p gains were often masked in the GGH profiles due to MYCN amplification. The commonly overrepresented region was chromosome segment 2pter-2p22, which contains the MYCN gene, and five out of eleven 2p breakpoints clustered to the interface of chromosome bands 2p16 and 2p21. In neuroblastoma cell line SJNB-12, with double minutes (dmins) but no MYCN amplification, the dmins were shown to be derived from 16q22-q23 sequences. The ATBF1 gene, an AT-binding transcription factor involved in normal neurogenesis and located at 16q22.2, was shown to be present in the amplicon. This is the first report describing the possible implication of ATBF1 in neuroblastoma cells. We conclude that a combined approach of M-FISH, cytogenetics, and CGH allowed a more complete and accurate description of the genetic alterations occurring in the investigated cell lines.
authors.affiliation: Centre for Medical Genetics, Ghent University Hospital, De Pintelaan 185, 9000 Ghent, Belgium.
authors.forename: N
authors.initials: N
authors.lastname: Van Roy
authors.forename: H
authors.initials: H
authors.lastname: Van Limbergen
authors.forename: J
authors.initials: J
authors.lastname: Vandesompele
authors.forename: M
authors.initials: M
authors.lastname: Van Gele
authors.forename: B
authors.initials: B
authors.lastname: Poppe
authors.forename: H
authors.initials: H
authors.lastname: Salwen
authors.forename: G
authors.initials: G
authors.lastname: Laureys
authors.forename: N
authors.initials: N
authors.lastname: Manoel
authors.forename: A
authors.initials: A
authors.lastname: De Paepe
authors.forename: F
authors.initials: F
authors.lastname: Speleman
mesh_terms: D020223:Chromosome Painting / Q000379:methods*; D005260:Female; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007621:Karyotyping; D008297:Male; D009447:Neuroblastoma / Q000235:genetics*; D009693:Nucleic Acid Hybridization / Q000379:methods; D014407:Tumor Cells, Cultured