Chromosoma

, Volume 115, Issue 6, pp 459–467 | Cite as

The breakage–fusion–bridge (BFB) cycle as a mechanism for generating genetic heterogeneity in osteosarcoma

  • Shamini Selvarajah
  • Maisa Yoshimoto
  • Paul C. Park
  • Georges Maire
  • Jana Paderova
  • Jane Bayani
  • Gloria Lim
  • Khaldoun Al-Romaih
  • Jeremy A. Squire
  • Maria Zielenska
Research article

Abstract

Osteosarcoma (OS) is characterized by chromosomal instability and high copy number gene amplification. The breakage–fusion–bridge (BFB) cycle is a well-established mechanism of genome instability in tumors and in vitro models used to study the origins of complex chromosomal rearrangements and cancer genome amplification. To determine whether the BFB cycle could be increasing the de novo rate of formation of cytogenetic aberrations in OS, the frequency of anaphase bridge configurations and dicentric chromosomes in four OS cell lines was quantified. An increased level of anaphase bridges and dicentrics was observed in all the OS cell lines. There was also a strong association between the frequencies of anaphase bridges, dicentrics, centrosomal anomalies, and multipolar mitotic figures in all the OS cell lines, indicating a possible link in the mechanisms that led to the structural and numerical instabilities observed in OS. In summary, this study has provided strong support for the role of the BFB cycle in generating the extensive structural chromosome aberrations, as well as cell-to-cell cytogenetic variation observed in OS, thus conferring the genetic diversity for OS tumor progression.

Notes

Acknowledgement

This work was supported by funding from the National Cancer Institute of Canada and Canadian Institute of Health Research (Ph.D. fellowship to S.S.).

References

  1. Akerman M, Dreinhofer K, Rydholm A, Willen H, Mertens F, Mitelman F, Mandahl N (1996) Cytogenetic studies on fine-needle aspiration samples from osteosarcoma and Ewing’s sarcoma. Diagn Cytopathol 15:17–22PubMedCrossRefGoogle Scholar
  2. Al-Romaih K, Bayani J, Vorobyova J, Karaskova J, Park PC, Zielenska M, Squire JA (2003) Chromosomal instability in osteosarcoma and its association with centrosome abnormalities. Cancer Genet Cytogenet 144:91–99PubMedCrossRefGoogle Scholar
  3. Atiye J, Wolf M, Kaur S, Monni O, Bohling T, Kivioja A, Tas E, Serra M, Tarkkanen M, Knuutila S (2005) Gene amplifications in osteosarcoma-CGH microarray analysis. Genes Chromosomes Cancer 42:158–163PubMedCrossRefGoogle Scholar
  4. Bayani J, Zielenska M, Pandita A, Al-Romaih K, Karaskova J, Harrison K, Bridge JA, Sorensen P, Thorner P, Squire JA (2003) Spectral karyotyping identifies recurrent complex rearrangements of chromosomes 8, 17, and 20 in osteosarcomas. Genes Chromosomes Cancer 36:7–16PubMedCrossRefGoogle Scholar
  5. Bosco G, Haber JE (1998) Chromosome break-induced DNA replication leads to nonreciprocal translocations and telomere capture. Genetics 150:1037–1047PubMedGoogle Scholar
  6. Cahill DP, Kinzler KW, Vogelstein B, Lengauer C (1999) Genetic instability and darwinian selection in tumours. Trends Cell Biol 9:M57–60PubMedCrossRefGoogle Scholar
  7. Eichler EE (1998) Masquerading repeats: paralogous pitfalls of the human genome. Genome Res 8:758–762PubMedGoogle Scholar
  8. Ghadimi BM, Sackett DL, Difilippantonio MJ, Schrock E, Neumann T, Jauho A, Auer G, Ried T (2000) Centrosome amplification and instability occurs exclusively in aneuploid, but not in diploid colorectal cancer cell lines, and correlates with numerical chromosomal aberrations. Genes Chromosomes Cancer 27:183–190PubMedCrossRefGoogle Scholar
  9. Gisselsson D (2003) Chromosome instability in cancer: how, when, and why? Adv Cancer Res 87:1–29PubMedCrossRefGoogle Scholar
  10. Gisselsson D, Hoglund M, Mertens F, Mandahl N (1999) Variable stability of chromosomes containing amplified alpha-satellite sequences in human mesenchymal tumours. Chromosoma 108:271–277PubMedCrossRefGoogle Scholar
  11. Gisselsson D, Pettersson L, Hoglund M, Heidenblad M, Gorunova L, Wiegant J, Mertens F, Dal Cin P, Mitelman F, Mandahl N (2000) Chromosomal breakage–fusion–bridge events cause genetic intratumor heterogeneity. Proc Natl Acad Sci USA 97:5357–5362PubMedCrossRefGoogle Scholar
  12. Gisselsson D, Jonson T, Yu C, Martins C, Mandahl N, Wiegant J, Jin Y, Mertens F, Jin C (2002) Centrosomal abnormalities, multipolar mitoses, and chromosomal instability in head and neck tumours with dysfunctional telomeres. Br J Cancer 87:202–207PubMedCrossRefGoogle Scholar
  13. Gisselsson D, Palsson E, Yu C, Mertens F, Mandahl N (2004) Mitotic instability associated with late genomic changes in bone and soft tissue tumours. Cancer Lett 206:69–76PubMedCrossRefGoogle Scholar
  14. Gollin SM (2004) Chromosomal instability. Curr Opin Oncol 16:25–31PubMedCrossRefGoogle Scholar
  15. Hoffelder DR, Luo L, Burke NA, Watkins SC, Gollin SM, Saunders WS (2004) Resolution of anaphase bridges in cancer cells. Chromosoma 112:389–397PubMedCrossRefGoogle Scholar
  16. Jin Y, Jin C, Wennerberg J, Hoglund M, Mertens F (2001) Cytogenetic and fluorescence in situ hybridization characterization of chromosome 8 rearrangements in head and neck squamous cell carcinomas. Cancer Genet Cytogenet 130:111–117PubMedCrossRefGoogle Scholar
  17. Kaufman RJ, Sharp PA, Latt SA (1983) Evolution of chromosomal regions containing transfected and amplified dihydrofolate reductase sequences. Mol Cell Biol 3:699–711PubMedGoogle Scholar
  18. Lim G, Karaskova J, Vukovic B, Bayani J, Beheshti B, Bernardini M, Squire JA, Zielenska M (2004) Combined spectral karyotyping, multicolor banding, and microarray comparative genomic hybridization analysis provides a detailed characterization of complex structural chromosomal rearrangements associated with gene amplification in the osteosarcoma cell line MG-63. Cancer Genet Cytogenet 153:158–164PubMedCrossRefGoogle Scholar
  19. Lim G, Karaskova J, Beheshti B, Vukovic B, Bayani J, Selvarajah S, Watson SK, Lam WL, Zielenska M, Squire JA (2005) An integrated mBAND and submegabase resolution tiling set (SMRT) CGH array analysis of focal amplification, microdeletions, and ladder structures consistent with breakage–fusion–bridge cycle events in osteosarcoma. Genes Chromosomes Cancer 42:392–403PubMedCrossRefGoogle Scholar
  20. Lo AW, Sprung CN, Fouladi B, Pedram M, Sabatier L, Ricoul M, Reynolds GE, Murnane JP (2002) Chromosome instability as a result of double-strand breaks near telomeres in mouse embryonic stem cells. Mol Cell Biol 22:4836–4850PubMedCrossRefGoogle Scholar
  21. Loeb LA (1991) Mutator phenotype may be required for multistage carcinogenesis. Cancer Res 51:3075–3079PubMedGoogle Scholar
  22. Masuda A, Takahashi T (2002) Chromosome instability in human lung cancers: possible underlying mechanisms and potential consequences in the pathogenesis. Oncogene 21:6884–6897PubMedCrossRefGoogle Scholar
  23. McClintock B (1941) The stability of broken ends of chromosomes in Zea mays. Genetics 26:234–282Google Scholar
  24. Mertens F, Mandahl N, Orndal C, Baldetorp B, Bauer HC, Rydholm A, Wiebe T, Willen H, Akerman M, Heim S et al (1993) Cytogenetic findings in 33 osteosarcomas. Int J Cancer 55:44–50PubMedGoogle Scholar
  25. Michor F, Iwasa Y, Vogelstein B, Lengauer C, Nowak MA (2005) Can chromosomal instability initiate tumorigenesis? Semin Cancer Biol 15:43–49PubMedCrossRefGoogle Scholar
  26. Mitelman F (1995) An International System for Human Cytogenetic Nomenclature (ISCN 1995). S. Karger, BaselGoogle Scholar
  27. Mussman JG, Horn HF, Carroll PE, Okuda M, Tarapore P, Donehower LA, Fukasawa K (2000) Synergistic induction of centrosome hyperamplification by loss of p53 and cyclin E overexpression. Oncogene 19:1635–1646PubMedCrossRefGoogle Scholar
  28. Nowak MA, Komarova NL, Sengupta A, Jallepalli PV, Shih Ie M, Vogelstein B, Lengauer C (2002) The role of chromosomal instability in tumor initiation. Proc Natl Acad Sci USA 99:16226–16231PubMedCrossRefGoogle Scholar
  29. Pfeiffer P, Goedecke W, Kuhfittig-Kulle S, Obe G (2004) Pathways of DNA double-strand break repair and their impact on the prevention and formation of chromosomal aberrations. Cytogenet Genome Res 104:7–13PubMedCrossRefGoogle Scholar
  30. Reshmi SC, Saunders WS, Kudla DM, Ragin CR, Gollin SM (2004) Chromosomal instability and marker chromosome evolution in oral squamous cell carcinoma. Genes Chromosomes Cancer 41:38–46PubMedCrossRefGoogle Scholar
  31. Saunders W (2005) Centrosomal amplification and spindle multipolarity in cancer cells. Semin Cancer Biol 15:25–32PubMedCrossRefGoogle Scholar
  32. Scheel C, Schaefer KL, Jauch A, Keller M, Wai D, Brinkschmidt C, van Valen F, Boecker W, Dockhorn-Dworniczak B, Poremba C (2001) Alternative lengthening of telomeres is associated with chromosomal instability in osteosarcomas. Oncogene 20:3835–3844PubMedCrossRefGoogle Scholar
  33. Shimizu N, Shingaki K, Kaneko-Sasaguri Y, Hashizume T, Kanda T (2005) When, where and how the bridge breaks: anaphase bridge breakage plays a crucial role in gene amplification and HSR generation. Exp Cell Res 302:233–243PubMedCrossRefGoogle Scholar
  34. Squire JA, Pei J, Marrano P, Beheshti B, Bayani J, Lim G, Moldovan L, Zielenska M (2003) High-resolution mapping of amplifications and deletions in pediatric osteosarcoma by use of CGH analysis of cDNA microarrays. Genes Chromosomes Cancer 38:215–225PubMedCrossRefGoogle Scholar
  35. Stewenius Y, Gorunova L, Jonson T, Larsson N, Hoglund M, Mandahl N, Mertens F, Mitelman F, Gisselsson D (2005) Structural and numerical chromosome changes in colon cancer develop through telomere-mediated anaphase bridges, not through mitotic multipolarity. Proc Natl Acad Sci USA 102:5541–5546PubMedCrossRefGoogle Scholar
  36. Stock C, Kager L, Fink FM, Gadner H, Ambros PF (2000) Chromosomal regions involved in the pathogenesis of osteosarcomas. Genes Chromosomes Cancer 28:329–336PubMedCrossRefGoogle Scholar
  37. Tarkkanen M, Karhu R, Kallioniemi A, Elomaa I, Kivioja AH, Nevalainen J, Bohling T, Karaharju E, Hyytinen E, Knuutila S et al (1995) Gains and losses of DNA sequences in osteosarcomas by comparative genomic hybridization. Cancer Res 55:1334–1338PubMedGoogle Scholar
  38. Toledo F, Le Roscouet D, Buttin G, Debatisse M (1992) Co-amplified markers alternate in megabase long chromosomal inverted repeats and cluster independently in interphase nuclei at early steps of mammalian gene amplification. EMBO J 11:2665–2673PubMedGoogle Scholar
  39. Tomescu O, Barr FG (2001) Chromosomal translocations in sarcomas: prospects for therapy. Trends Mol Med 7:554–559PubMedCrossRefGoogle Scholar
  40. Trask BJ, Hamlin JL (1989) Early dihydrofolate reductase gene amplification events in CHO cells usually occur on the same chromosome arm as the original locus. Genes Dev 3:1913–1925PubMedGoogle Scholar
  41. Tsuchiya T, Sekine K, Hinohara S, Namiki T, Nobori T, Kaneko Y (2000) Analysis of the p16INK4, p14ARF, p15, TP53, and MDM2 genes and their prognostic implications in osteosarcoma and Ewing sarcoma. Cancer Genet Cytogenet 120:91–98PubMedCrossRefGoogle Scholar
  42. Van Hooser AA, Ouspenski II, Gregson HC, Starr DA, Yen TJ, Goldberg ML, Yokomori K, Earnshaw WC, Sullivan KF, Brinkley BR (2001) Specification of kinetochore-forming chromatin by the histone H3 variant CENP-A. J Cell Sci 114:3529–3542PubMedGoogle Scholar
  43. Wang Y, Putnam CD, Kane MF, Zhang W, Edelmann L, Russell R, Carrion DV, Chin L, Kucherlapati R, Kolodner RD, Edelmann W (2005) Mutation in Rpa1 results in defective DNA double-strand break repair, chromosomal instability and cancer in mice. Nat Genet 37:750–755PubMedCrossRefGoogle Scholar
  44. Warburton PE, Cooke CA, Bourassa S, Vafa O, Sullivan BA, Stetten G, Gimelli G, Warburton D, Tyler-Smith C, Sullivan KF, Poirier GG, Earnshaw WC (1997) Immunolocalization of CENP-A suggests a distinct nucleosome structure at the inner kinetochore plate of active centromeres. Curr Biol 7:901–904PubMedCrossRefGoogle Scholar
  45. Zhu C, Mills KD, Ferguson DO, Lee C, Manis J, Fleming J, Gao Y, Morton CC, Alt FW (2002) Unrepaired DNA breaks in p53-deficient cells lead to oncogenic gene amplification subsequent to translocations. Cell 109:811–821PubMedCrossRefGoogle Scholar
  46. Zielenska M, Bayani J, Pandita A, Toledo S, Marrano P, Andrade J, Petrilli A, Thorner P, Sorensen P, Squire JA (2001) Comparative genomic hybridization analysis identifies gains of 1p35 approximately p36 and chromosome 19 in osteosarcoma. Cancer Genet Cytogenet 130:14–21PubMedCrossRefGoogle Scholar
  47. Zielenska M, Marrano P, Thorner P, Pei J, Beheshti B, Ho M, Bayani J, Liu Y, Sun BC, Squire JA, Hao XS (2004) High-resolution cDNA microarray CGH mapping of genomic imbalances in osteosarcoma using formalin-fixed paraffin-embedded tissue. Cytogenet Genome Res 107:77–82PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Shamini Selvarajah
    • 1
    • 2
  • Maisa Yoshimoto
    • 3
  • Paul C. Park
    • 4
  • Georges Maire
    • 3
  • Jana Paderova
    • 3
  • Jane Bayani
    • 3
  • Gloria Lim
    • 1
  • Khaldoun Al-Romaih
    • 2
    • 3
  • Jeremy A. Squire
    • 2
    • 3
    • 5
  • Maria Zielenska
    • 1
    • 2
  1. 1.Department of Pathology and Laboratory MedicineThe Hospital for Sick ChildrenTorontoCanada
  2. 2.Department of Laboratory Medicine and PathobiologyUniversity of TorontoTorontoCanada
  3. 3.Applied Molecular Oncology, Ontario Cancer InstitutePrincess Margaret HospitalTorontoCanada
  4. 4.Department of Pathology and Laboratory Medicine and PathobiologyOttawa General HospitalOttawaCanada
  5. 5.Department of Medical Biophysics, Faculty of MedicineUniversity of TorontoTorontoCanada

Personalised recommendations