Skip to main content
SpringerLink
Log in

Proliferation of aneuploid cells induced by CENP-E depletion is counteracted by the p14ARF tumor suppressor

  • Original Article
  • Published:
Molecular Genetics and Genomics Aims and scope Submit manuscript

Abstract

The spindle assembly checkpoint (SAC) is a cellular surveillance mechanism that ensures the fidelity of chromosomes segregation. Reduced expression of some of its components weakens the SAC and induces chromosome instability and aneuploidy, which are both well-known hallmarks of cancer cells. Centromere protein-E (CENP-E) is a crucial component of the SAC and its function is to facilitate kinetochore microtubule attachment required to achieve and maintain chromosome alignment. The present study investigates the possible role of p14ARF as a controller of aneuploid cells proliferation. We used RNA interference to induce aneuploidy by partial depletion of CENP-E in human primary fibroblasts (IMR90) and in near diploid tumor cells (HCT116). In contrast to IMR90 aneuploid cell number, which was drastically reduced and leaned towards the WT condition, HCT116 aneuploid cell numbers were slightly decreased at later time points. This euploidy restoration was accompanied by increased p14ARF expression in IMR90 cells and followed ectopic p14ARF re-expression in p14ARF-null HCT116 cells. Collectively, our results suggest that hampering proliferation of aneuploid cells could be an additional role of the p14ARF tumor suppressor.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Abrieu A, Kahana JA, Wood KW, Cleveland DW (2000) CENP-E as an essential component of the mitotic checkpoint in vitro. Cell 102:817–826

    Article  CAS  PubMed  Google Scholar 

  • Ayrault O, Andrique L, Fauvin D, Eymin B, Gazzeri S, Séité P (2006) Human tumor suppressor p14ARF negatively regulates rRNA transcription and inhibits UBF1 transcription factor phosphorylation. Oncogene 25:7577–7586

    Article  CAS  PubMed  Google Scholar 

  • Bakhoum SF, Compton DA (2012) Chromosomal instability and cancer: a complex relationship with therapeutic potential. J Clin Invest 122:1138–1143

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Barra V, Schillaci T, Lentini L, Costa G, Di Leonardo A (2012) Bypass of cell cycle arrest induced by transient DNMT1 post-transcriptional silencing triggers aneuploidy in human cells. Cell Div 7:2

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bennett A, Bechi B, Tighe A, Thompson S, Procter DJ, Taylor SS (2015) Cenp-E inhibitor GSK923295: Novel synthetic route and use as a tool to generate aneuploidy. Oncotarget 6:20921–20932

    Article  PubMed  PubMed Central  Google Scholar 

  • Britigan EMC, Wan J, Zasadil LM, Ryan SD, Weaver BA (2014) The ARF tumor suppressor prevents chromosomal instability and ensures mitotic checkpoint fidelity through regulation of Aurora B. Mol Biol Cell 25:2761–2773

    Article  PubMed  PubMed Central  Google Scholar 

  • Burri N, Shaw P, Bouzourene H, Sordat I, Sordat B, Gillet M, Schorderet D, Bosman FT, Chaubert P (2001) Methylation silencing and mutations of the p14ARF and p16INK4a genes in colon cancer. Lab Invest 81:217–229

    Article  CAS  PubMed  Google Scholar 

  • Ganmore I, Smooha G, Izraeli S (2009) Constitutional aneuploidy and cancer predisposition. Hum Mol Genet 18:84–93

    Article  CAS  Google Scholar 

  • Giam M, Rancati G (2015) Aneuploidy and chromosomal instability in cancer: a jackpot to chaos. Cell Div 10:3

    Article  PubMed  PubMed Central  Google Scholar 

  • Gordon DJ, Resio B, Pellman D (2012) Causes and consequences of aneuploidy in cancer. Nat Rev Genet 13:189–203

    Article  CAS  PubMed  Google Scholar 

  • Holland AJ, Cleveland DW (2009) Boveri revisited: chromosomal instability, aneuploidy and tumorigenesis. Nat Rev Mol Cell Biol 10:478–487

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Holland AJ, Cleveland DW (2012) Losing balance: the origin and impact of aneuploidy in cancer. EMBO Rep 13:501–514

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Janssen A, van der Burg M, Szuhai K, Kops GJ, Medema RH (2011) Chromosome segregation errors as a cause of DNA damage and structural chromosome aberrations. Science 333:1895–1898

    Article  CAS  PubMed  Google Scholar 

  • Lengauer C, Kinzler KW, Vogelstein B (1997) Genetic instability in colorectal cancers. Nature 386:623–627

    Article  CAS  PubMed  Google Scholar 

  • Lentini L, Amato A, Schillaci T, Di Leonardo A (2007) Simultaneous aurora-A/STK15 overexpression and centrosome amplification induce chromosomal instability in tumour cells with a MIN phenotype. BMC Cancer 7:212

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lentini L, Barra V, Schillaci T, Di Leonardo A (2012) MAD2 depletion triggers premature cellular senescence in human primary fibroblasts by activating a P53 pathway preventing aneuploid cells propagation. J Cell Physiol 227:3324–3332

    Article  CAS  PubMed  Google Scholar 

  • Li M, Fang X, Baker DJ, Guo L, Gao X, Wei Z, Han S, van Deursen JM, Zhang P (2010) The ATM – p53 pathway suppresses aneuploidy- induced tumorigenesis. Proc Natl Acad Sci U S A 107(32):14188–14193

    Article  PubMed  PubMed Central  Google Scholar 

  • Ohashi A, Ohori M, Iwai K, Nambu T, Miyamoto M, Kawamoto T, Okaniwa M (2015) A novel time-dependent CENP-E inhibitor with potent antitumor activity. PLoS One 10(12):e0144675

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Orr B, Godek KM, Compton D (2015) Aneuploidy. Curr Biol 25:R538–R542

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pear W, Nolan G, Scott M (1993) Production of high-titer helper-free retroviruses by transient transfection. Proc Natl Acad Sci U S A 90(18):8392–8396

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Schaar BT, Chan GK, Maddox P, Salmon ED, Yen TJ (1997)) CENP-E function at kinetochores is essential for chromosome alignment. J Cell Biol 139:1373–1382

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Siegel JJ, Amon A (2012) New insights into the troubles of aneuploidy. Annu Rev Cell Dev Biol 28:189–214

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Silk AD, Zasadil LM, Holland AJ, Vitre B, Cleveland DW, Weaver B (2013) Chromosome missegregation rate predicts whether aneuploidy will promote or suppress tumors. Proc Natl Acad Sci USA 110, E4134–41

    Article  Google Scholar 

  • Soto M, Raaijmakers JA, Bakker B, Spierings DCJ, Lansdorp PM, Foijer F, Medema RH (2017) p53 prohibits propagation of chromosome segregation errors that produce structural aneuploidies. Cell Rep 19(12):2423–2431

    Article  CAS  PubMed  Google Scholar 

  • Thompson SL, Compton D (2010) Proliferation of aneuploid human cells is limited by a p53-dependent mechanism. J Cell Biol 188:369–381

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Thompson SL, Compton DA (2011) Chromosomes and cancer cells. Chromosom Res 19:433–444

    Article  Google Scholar 

  • Thompson SL, Bakhoum SF, Compton DA (2010) Mechanisms of chromosomal instability. Curr Biol 20:R285–R295

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Torres EM, Sokolsky T, Tucker CM, Chan LY, Boselli M, Dunham MJ, Amon A (2007) Effects of aneuploidy on cellular physiology and cell division in haploid yeast. Science 317:916–924

    Article  CAS  PubMed  Google Scholar 

  • Torres EM, Williams BR, Amon A (2008) Aneuploidy: cells losing their balance. Genetics 179, 737–746

    Article  CAS  Google Scholar 

  • Veneziano L, Barra V, Lentini L, Spatafora S, Di Leonardo A (2016) p14ARF prevents proliferation of aneuploid cells by inducing p53-dependent apoptosis. J Cell Physiol 231:336–344

    Article  CAS  PubMed  Google Scholar 

  • Weaver BA, Cleveland DW (2006) Does aneuploidy cause cancer? Curr Opin Cell Biol 18:658–667

    Article  CAS  PubMed  Google Scholar 

  • Weaver BA, Cleveland DW (2007)Aneuploidy: instigator and inhibitor of tumorigenesis. Cancer Res 67:10103–10105

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Weaver BA, Bonday ZQ, Putkey FR, Kops GJPL, Silk AD, Cleveland DW (2003) Centromere-associated protein-E is essential for the mammalian mitotic checkpoint to prevent aneuploidy due to single chromosome loss. J Cell Biol 162:551–563

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Weaver BA, Silk AD, Montagna C, Verdier-Pinard P, Cleveland DW (2007) Aneuploidy acts both oncogenically and as a tumor suppressor. Cancer Cell 11:25–36

    Article  CAS  PubMed  Google Scholar 

  • Yen TJ, Li G, Schaar BT, Szilak I, Cleveland DW (1992) CENP-E is a putative kinetochore motor that accumulates just before mitosis. Nature 359:536–539

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was in part supported by Università degli Studi di Palermo, Italy: ATE-0255, and by Centro di Oncobiologia Sperimentale (COBS) to ADL. We want to thank Prof. S. Gazzeri, University J. Fourier, La Tronche, France, for providing us with the pcDNA3.1 plasmid harboring the p14ARF c-DNA. We are grateful to Prof. Floris Foijer, European Research Institute for the Biology of Ageing (ERIBA) University Medical Center Groningen (the Netherlands) and to Dr. Rosario Billetta, Abilita Bio, Inc. San Diego, CA (United States of America) and to Maha Mohammad Said, Institut Gustave Roussy, Villejuif (France) for their thoughtful reading of the manuscript.

Author information

Author notes

  1. Viviana Barra

    Present address: Department of Genetic Stability and Oncogenesis, Institut Gustave Roussy, CNRS UMR8200, 94805, Villejuif, France

  2. Lorena Veneziano and Viviana Barra contributed equally to this study.

Authors and Affiliations

  1. Department of Biological, Chemical and Pharmaceutical Sciences and Technologies, University of Palermo, Palermo, Italy

    Lorena Veneziano, Viviana Barra, Danilo Cilluffo & Aldo Di Leonardo

  2. Centro di OncoBiologia Sperimentale (COBS), Palermo, Italy

    Aldo Di Leonardo

Authors
  1. Lorena Veneziano
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Viviana Barra
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Danilo Cilluffo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Aldo Di Leonardo
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

LV did RNAi experiments, cell transfection, cytogenetics analysis, vector construction, RT-qPCR, analysed the data and helped to write the manuscript; VB did RNAi, cell transfection, cytogenetics analysis, RT-qPCR, analysed the data and helped to write the manuscript; DC did immunofluorescence microscopy, Western Blot and helped with vector construction; ADL designed the experiments, analysed the data and wrote the manuscript.

Corresponding author

Correspondence to Aldo Di Leonardo.

Ethics declarations

Conflict of interest

The authors declare that they have no competing interests.

Additional information

Communicated by Andres Aguilera.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Veneziano, L., Barra, V., Cilluffo, D. et al. Proliferation of aneuploid cells induced by CENP-E depletion is counteracted by the p14ARF tumor suppressor. Mol Genet Genomics 294, 149–158 (2019). https://doi.org/10.1007/s00438-018-1495-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00438-018-1495-5

Keywords

Search

Navigation