Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Chromosome nondisjunction yields tetraploid rather than aneuploid cells in human cell lines

Abstract

Although mutations in cell cycle regulators or spindle proteins can perturb chromosome segregation1,2,3,4,5,6,7, the causes and consequences of spontaneous mitotic chromosome nondisjunction in human cells are not well understood. It has been assumed that nondisjunction of a chromosome during mitosis will yield two aneuploid daughter cells. Here we show that chromosome nondisjunction is tightly coupled to regulation of cytokinesis in human cell lines, such that nondisjunction results in the formation of tetraploid rather than aneuploid cells. We observed that spontaneously arising binucleated cells exhibited chromosome mis-segregation rates up to 166-fold higher than the overall mitotic population. Long-term imaging experiments indicated that most binucleated cells arose through a bipolar mitosis followed by regression of the cleavage furrow hours later. Nondisjunction occurred with high frequency in cells that became binucleated by furrow regression, but not in cells that completed cytokinesis to form two mononucleated cells. Our findings indicate that nondisjunction does not directly yield aneuploid cells, but rather tetraploid cells that may subsequently become aneuploid through further division. The coupling of spontaneous segregation errors to furrow regression provides a potential explanation for the prevalence of hyperdiploid chromosome number and centrosome amplification observed in many cancers8,9.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Chromosome mis-segregation occurs at high frequency in spontaneously arising binucleated cells.
Figure 2: Nondisjunction occurs with high frequency in cells that become binucleated by furrow regression, but not in cells that complete cytokinesis.
Figure 3: Fates of mononucleated and binucleated N/TERT-1 and HeLa cells determined by long-term imaging.
Figure 4: Model summarizing the relationship of chromosome mis-segregation in the regulation of cytokinesis, and subsequent possible fates of resulting binucleated cells.

References

  1. Cahill, D. P. et al. Mutations of mitotic checkpoint genes in human cancers. Nature 392, 300–303 (1998)

    ADS  CAS  Article  Google Scholar 

  2. Fodde, R. et al. Mutations in the APC tumour suppressor gene cause chromosomal instability. Nature Cell Biol. 3, 433–438 (2001)

    CAS  Article  Google Scholar 

  3. Hernando, E. et al. Rb inactivation promotes genomic instability by uncoupling cell cycle progression from mitotic control. Nature 430, 797–802 (2004)

    ADS  CAS  Article  Google Scholar 

  4. Jallepalli, P. V. et al. Securin is required for chromosomal stability in human cells. Cell 105, 445–457 (2001)

    CAS  Article  Google Scholar 

  5. Kaplan, K. B. et al. A role for the Adenomatous Polyposis Coli protein in chromosome segregation. Nature Cell Biol. 3, 429–432 (2001)

    CAS  Article  Google Scholar 

  6. Meraldi, P., Honda, R. & Nigg, E. A. Aurora-A overexpression reveals tetraploidization as a major route to centrosome amplification in p53-/- cells. EMBO J. 21, 483–492 (2002)

    CAS  Article  Google Scholar 

  7. Rajagopalan, H. et al. Inactivation of hCDC4 can cause chromosomal instability. Nature 428, 77–81 (2004)

    ADS  CAS  Article  Google Scholar 

  8. Nigg, E. A. Centrosome aberrations: cause or consequence of cancer progression? Nature Rev. Cancer 2, 815–825 (2002)

    CAS  Article  Google Scholar 

  9. Storchova, Z. & Pellman, D. From polyploidy to aneuploidy, genome instability and cancer. Nature Rev. Mol. Cell Biol. 5, 45–54 (2004)

    CAS  Article  Google Scholar 

  10. Bharadwaj, R. & Yu, H. The spindle checkpoint, aneuploidy and cancer. Oncogene 23, 2016–2027 (2004)

    CAS  Article  Google Scholar 

  11. Draviam, V. M., Xie, S. & Sorger, P. K. Chromosome segregation and genomic stability. Curr. Opin. Genet. Dev. 14, 120–125 (2004)

    CAS  Article  Google Scholar 

  12. Carere, A. et al. Analysis of chromosome loss and non-disjunction in cytokinesis-blocked lymphocytes of 24 male subjects. Mutagenesis 14, 491–496 (1999)

    CAS  Article  Google Scholar 

  13. Cimini, D., Tanzarella, C. & Degrassi, F. Differences in malsegregation rates obtained by scoring ana-telophases or binucleate cells. Mutagenesis 14, 563–568 (1999)

    CAS  Article  Google Scholar 

  14. Dickson, M. A. et al. Human keratinocytes that express hTERT and also bypass a p16(INK4a)-enforced mechanism that limits life span become immortal yet retain normal growth and differentiation characteristics. Mol. Cell. Biol. 20, 1436–1447 (2000)

    CAS  Article  Google Scholar 

  15. Macville, M. et al. Comprehensive and definitive molecular cytogenetic characterization of HeLa cells by spectral karyotyping. Cancer Res. 59, 141–150 (1999)

    CAS  PubMed  Google Scholar 

  16. Berger, R. et al. Androgen-induced differentiation and tumorigenicity of human prostate epithelial cells. Cancer Res. 64, 8867–8875 (2004)

    CAS  Article  Google Scholar 

  17. Tanabe, K., Ikegami, Y., Ishida, R. & Andoh, T. Inhibition of topoisomerase II by antitumor agents bis(2,6-dioxopiperazine) derivatives. Cancer Res. 51, 4903–4908 (1991)

    CAS  PubMed  Google Scholar 

  18. Kanda, T., Sullivan, K. F. & Wahl, G. M. Histone-GFP fusion protein enables sensitive analysis of chromosome dynamics in living mammalian cells. Curr. Biol. 8, 377–385 (1998)

    CAS  Article  Google Scholar 

  19. Mullins, J. M. & Biesele, J. J. Terminal phase of cytokinesis in D-98s cells. J. Cell Biol. 73, 672–684 (1977)

    CAS  Article  Google Scholar 

  20. Cimini, D. et al. Merotelic kinetochore orientation is a major mechanism of aneuploidy in mitotic mammalian tissue cells. J. Cell Biol. 153, 517–527 (2001)

    CAS  Article  Google Scholar 

  21. Uetake, Y. & Sluder, G. Cell cycle progression after cleavage failure: mammalian somatic cells do not possess a “tetraploidy checkpoint”. J. Cell Biol. 165, 609–615 (2004)

    CAS  Article  Google Scholar 

  22. Wong, C. & Stearns, T. Mammalian cells lack checkpoints for tetraploidy, aberrant centrosome number, and cytokinesis failure. BMC Cell Biol. 6, 6 (2005)

    Article  Google Scholar 

  23. Kops, G. J., Foltz, D. R. & Cleveland, D. W. Lethality to human cancer cells through massive chromosome loss by inhibition of the mitotic checkpoint. Proc. Natl Acad. Sci. USA 101, 8699–8704 (2004)

    ADS  CAS  Article  Google Scholar 

  24. Galipeau, P. C. et al. 17p (p53) allelic losses, 4N (G2/tetraploid) populations, and progression to aneuploidy in Barrett's esophagus. Proc. Natl Acad. Sci. USA 93, 7081–7084 (1996)

    ADS  CAS  Article  Google Scholar 

  25. Burholt, D. R. et al. Karyotypic evolution of a human undifferentiated large cell carcinoma of the lung in tissue culture. Cancer Res. 49, 3355–3361 (1989)

    CAS  PubMed  Google Scholar 

  26. Kaneko, Y. & Knudson, A. G. Mechanism and relevance of ploidy in neuroblastoma. Genes Chromosom. Cancer 29, 89–95 (2000)

    CAS  Article  Google Scholar 

  27. Shackney, S. E. et al. Model for the genetic evolution of human solid tumors. Cancer Res. 49, 3344–3354 (1989)

    CAS  PubMed  Google Scholar 

  28. Duesberg, P. & Li, R. Multistep carcinogenesis: a chain reaction of aneuploidizations. Cell Cycle 2, 202–210 (2003)

    CAS  Article  Google Scholar 

  29. Mitelman, F., Johansson, B. & Mertens, F. (eds) Mitelman database of chromosome alterations in cancer. http://cgap.nci.nih.gov/Chromosomes/Mitelman (2005).

  30. Shi, Q. et al. Increased nondisjunction of chromosome 21 with age in human peripheral lymphocytes. Mutat. Res. 452, 27–36 (2000)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank J. Rheinwald for N/TERT-1 cells, W. Hahn for PrEC cells, J. Waters and the Nikon Imaging Center at Harvard Medical School for assistance and equipment, T. Mitchison for discussions, and D. Pellman, A. Amon, D. Moazed and P. Jackson for comments on the manuscript. This work was supported by the Harry C. McKenzie Family Foundation and the Harvard-Armenise Foundation. R.W.K. is a Damon Runyon-Walter Winchell Foundation Scholar.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Randall W. King.

Ethics declarations

Competing interests

Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Supplementary information

Supplementary Notes

This file contains Supplementary Methods, Supplementary Figures S1–S7, Supplementary Tables S1–S3, and legends for the Supplementary Videos. (PDF 2866 kb)

Supplementary Movie S1

This movie shows a mononucleated HeLa cell expressing an H2B–GFP fusion protein that undergoes a normal bipolar mitosis. (MOV 377 kb)

Supplementary Movie S2

This movie shows the generation of a binucleated cell through normal bipolar mitosis followed by cleavage furrow regression. (MOV 2040 kb)

Supplementary Movie S3

This movie shows a second example of the generation of a binucleated cell through normal bipolar mitosis followed by cleavage furrow regression. (MOV 1223 kb)

Supplementary Movie S4

This movie shows the generation of a binucleated cell through abnormal mitosis of a mononucleated cell. (MOV 934 kb)

Supplementary Movie S5

This movie shows the generation of a binucleated cell through fusion of two newly generated mononucleated cells. (MOV 1161 kb)

Supplementary Movie S6

This movie shows a binucleated cell that divides with a tetrapolar mitosis to produce two binucleated cells. (MOV 1063 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Shi, Q., King, R. Chromosome nondisjunction yields tetraploid rather than aneuploid cells in human cell lines. Nature 437, 1038–1042 (2005). https://doi.org/10.1038/nature03958

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature03958

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing