Spectral karyotyping identifies recurrent complex rearrangements of chromosomes 8, 17, and 20 in osteosarcomas

Department of Laboratory Medicine and Pathobiology, University of Toronto, Ontario, Canada.
Genes Chromosomes and Cancer (Impact Factor: 3.84). 01/2003; 36(1):7-16. DOI: 10.1002/gcc.10132
Source: PubMed

ABSTRACT Conventional cytogenetic studies have shown that osteosarcomas (OSs) are often highly aneuploid, with a large number of both structural and numerical chromosomal alterations. To investigate the complexity of OS karyotypes in detail, we applied spectral karyotyping (SKY) to a series of 14 primary OS tumors and four established OS cell lines. A total of 531 rearrangements were identified by SKY, of which 300 breakpoints could be assigned to a specific chromosome band. There was an average of 38.5 breakpoints identified by SKY per primary tumor. Chromosome 20 was involved in a disproportionately high number of structural rearrangements, with 38 different aberrations being detected. Chromosomal rearrangements between chromosomes 20 and 8 were evident in four tumors. FISH analysis using a 20q13 subtelomeric probe identified frequent involvement of 20q in complex structural rearrangements of OS cell lines. Characterization of the structural aberrations of chromosomes 8 and 17 by use of SKY demonstrated frequent duplication or partial gains of chromosome bands 8q23-24 and 17p11-13. Other chromosomes frequently involved in structural alteration were chromosomes 1 (47 rearrangements) and 6 (38 rearrangements). Centromeric rearrangements often involving chromosomes 1, 6, 13, 14, 17, and 20 were present. Four of the 14 primary OS tumors were characterized by nonclonal changes that included both structural and numerical alterations. In summary, OS tumors have a very high frequency of structural and numerical alterations, compounded by gross changes in ploidy. This intrinsic karyotype instability leads to a diversity of rearrangements and the acquisition of composite chromosomal rearrangements, with the highest frequency of alteration leading to gain of 8q23-24 and 17p11-13 and rearrangement of 20q. These findings suggest that specific sequences mapping to these chromosomal regions will likely have a role in the development and progression of OS.

  • Source
    • "Interestingly, AURK-B is located on chromosome 17p13.1, which has also been found to be amplified in OS (Bayani et al, 2003). This body of evidence has indicated the Aurora kinases as promising candidates to develop targeted drugs. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Background: Aurora kinases are key regulators of cell cycle and represent new promising therapeutic targets in several human tumours. Methods: Biological relevance of Aurora kinase-A and -B was assessed on osteosarcoma clinical samples and by silencing these genes with specific siRNA in three human osteosarcoma cell lines. In vitro efficacy of two Aurora kinases-targeting drugs (VX-680 and ZM447439) was evaluated on a panel of four drug-sensitive and six drug-resistant human osteosarcoma cell lines. Results: Human osteosarcoma cell lines proved to be highly sensitive to both drugs. A decreased drug sensitivity was observed in doxorubicin-resistant cell lines, most probably related to ABCB1/MDR1 overexpression. Both drugs variably induced hyperploidy and apoptosis in the majority of cell lines. VX-680 also reduced in vitro cell motility and soft-agar cloning efficiency. Drug association experiments showed that VX-680 positively interacts with all conventional drugs used in osteosarcoma chemotherapy, overcoming the cross-resistance observed in the single-drug treatments. Conclusion: Aurora kinase-A and -B represent new candidate therapeutic targets for osteosarcoma. In vitro analysis of the Aurora kinases inhibitors VX-680 and ZM447439 indicated in VX-680 a new promising drug of potential clinical usefulness in association with conventional osteosarcoma chemotherapeutic agents.
    British Journal of Cancer 10/2013; 109(10). DOI:10.1038/bjc.2013.643 · 4.82 Impact Factor
  • Source
    • "Such reorganization includes a high degree of aneuploidy, gene amplification , and multiple unbalanced chromosomal rearrangements (Bridge et al., 1997; Bayani et al., 2003; Ozaki et al., 2003; Lim et al., 2004; Man et al., 2004) Analysis of human OS by array comparative genomic hybridization (aCGH) has revealed high amplitude copy number gain (log 2 ratio of tumor:reference !1.0) or amplification (log 2 ratio of tumor:reference !2.0) to be frequent events in human OS, particularly within the cytogenetic regions defined by 6p22-p21, 8q24 and 17p12-p11.2 (Forus et al., 1995; Tarkkanen et al., 1995, 1998, 1999; Simons et al., 1997; Zielenska et al., 2001; Bayani et al., 2003; Squire et al., 2003; Selvarajah et al., 2008). The frequency of occurrence of recurrent genomic amplifications suggests that they may harbor genes important in tumorigenesis and/or tumor progression (Lu et al., 2008; Sadikovic et al., 2009). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Osteosarcoma (OS) is the most commonly diagnosed malignant bone tumor in humans and dogs, characterized in both species by extremely complex karyotypes exhibiting high frequencies of genomic imbalance. Evaluation of genomic signatures in human OS using array comparative genomic hybridization (aCGH) has assisted in uncovering genetic mechanisms that result in disease phenotype. Previous low-resolution (10-20 Mb) aCGH analysis of canine OS identified a wide range of recurrent DNA copy number aberrations, indicating extensive genomic instability. In this study, we profiled 123 canine OS tumors by 1 Mb-resolution aCGH to generate a dataset for direct comparison with current data for human OS, concluding that several high frequency aberrations in canine and human OS are orthologous. To ensure complete coverage of gene annotation, we identified the human refseq genes that map to these orthologous aberrant dog regions and found several candidate genes warranting evaluation for OS involvement. Specifically, subsequenct FISH and qRT-PCR analysis of RUNX2, TUSC3, and PTEN indicated that expression levels correlated with genomic copy number status, showcasing RUNX2 as an OS associated gene and TUSC3 as a possible tumor suppressor candidate. Together these data demonstrate the ability of genomic comparative oncology to identify genetic abberations which may be important for OS progression. Large scale screening of genomic imbalance in canine OS further validates the use of the dog as a suitable model for human cancers, supporting the idea that dysregulation discovered in canine cancers will provide an avenue for complementary study in human counterparts.
    Genes Chromosomes and Cancer 11/2011; 50(11):859-74. DOI:10.1002/gcc.20908 · 3.84 Impact Factor
  • Source
    • "Fluorescence spectral karyotyping (SKY) was performed on the metaphase cells according to the probe manufacturer's instructions (ASI, Carlsbad, CA) and as previously published [Bayani et al., 2003]. Spectral images were acquired with an SD200 spectral bioimaging system (Applied Spectral Imaging [ASI], Migdal Ha'Emek, Israel) and analyzed using the ASI SkyView software (version 1.2). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Supernumerary marker chromosomes (sSMC) may or may not be associated with an abnormal phenotype, depending on the presence of euchromatin, on their chromosomal origin and whether they are inherited. Over 80% of sSMCs are derived from acrocentric chromosomes and half of them include the short arm of chromosome 15. Generally, they appear as bisatellited isodicentric marker chromosomes, most of them are symmetric. These chromosomes are normally originated de novo and are associated with mild to severe intellectual disability but not with physical abnormalities. We report on a patient with an SMC studied using classical and molecular cytogenetic procedures (G and C banding, NOR staining, painting and centromeric fluorescent in situ hybridization (FISH), BAC-FISH, and SKY). The MLPA technique and DNA polymorphic markers were used in order to identify its parental origin. The marker chromosome, monosatellited and monocentric, was found to be derived from a maternal chromosome 15 and was defined as 15pter-q21.2. This is the report of the largest de novo monosatellited 15q marker chromosome ever published presenting detailed cytogenetic and clinical data. It was associated with a phenotype including cardiac defect, absence of septum pellucidum, and dysplasia of the corpus callosum.
    American Journal of Medical Genetics Part A 03/2010; 152A(3):753-8. DOI:10.1002/ajmg.a.33308 · 2.05 Impact Factor
Show more