Isolation of osteosarcoma-associated amplified DNA sequences using representational difference analysis.
Comparative genomic hybridization analysis of a primary osteosarcoma and its metastasis revealed two regions of DNA amplification, one at 17p11.2-12 and one at 19q12-13. Subsequent representational difference analysis of the primary tumor resulted in the isolation of two distinct tumor-amplified DNA fragments originating from chromosome 19. A YAC clone corresponding to one of the two isolated DNA fragments was used for fluorescence in situ hybridization on normal human lymphocyte metaphases and tumor-derived nuclei. This resulted in the localization of this YAC to 19q12-13.1 and confirmed the amplification status of the isolated fragment in the tumors. The availability of such RDA-isolated sequences may be instrumental in the search for genes relevant for tumor development.
Simons
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Department of Human Genetics, University Hospital, Nijmegen, The Netherlands. A.Simons@antrg.azn.nl
Janssen
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Suijkerbuijk
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RF
Veth
R P
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Pruszczynski
M
M
Hulsbergen-van de Kaa
C A
CA
du Manoir
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S
Geurts van Kessel
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D000293:Adolescent; D015139:Blotting, Southern; D001859:Bone Neoplasms / Q000235:genetics* / Q000473:pathology; D002874:Chromosome Mapping; D018244:Chromosomes, Artificial, Yeast; D002886:Chromosomes, Human, Pair 17 / Q000235:genetics; D002888:Chromosomes, Human, Pair 19 / Q000235:genetics; D004273:DNA, Neoplasm / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D005260:Female; D005784:Gene Amplification / Q000235:genetics*; D005819:Genetic Markers; D015698:Genomic Library; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D012516:Osteosarcoma / Q000235:genetics* / Q000556:secondary; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D004273:DNA, Neoplasm; D005819:Genetic Markers