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Senescence bypass screen identifies TBX2, which represses Cdkn2a (p19ARF) and is amplified in a subset of human breast cancers

Nature Genetics volume 26, pages 291–299 (2000)Cite this article

Abstract

To identify new immortalizing genes with potential roles in tumorigenesis, we performed a genetic screen aimed to bypass the rapid and tight senescence arrest of primary fibroblasts deficient for the oncogene Bmi1. We identified the T-box member TBX2 as a potent immortalizing gene that acts by downregulating Cdkn2a (p19ARF). TBX2 represses the Cdkn2a (p19ARF) promoter and attenuates E2F1, Myc or HRAS-mediated induction of Cdkn2a (p19ARF). We found TBX2 to be amplified in a subset of primary human breast cancers, indicating that it might contribute to breast cancer development.

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Figure 1: Schematic outline of the experimental set-up for rescue of Bmi1−/− MEFs using retroviral cDNA libraries.
Figure 2: Results of a secondary screen in which Bmi1−/− MEFs were infected with MoMuLV-mobilized control (empty) virus (a), mobilized Bmi1 virus (b), virus from a spontaneously immortalized primary clone (c), or virus from a TBX2 immortalized primary clone (d).
Figure 3: TBX2 immortalizes MEFs and delays senescence of primary human fibroblasts.
Figure 4: TBX2 downregulates Cdkn2a.
Figure 5: TBX2 overexpression leaves pathways downstream of Cdkn2a intact.
Figure 6: TBX2 counteracts induction of Cdkn2a (p19ARF) by Myc and HRAS.
Figure 7: TBX2 represses the CDKN2A (ARF) promoter.
Figure 8: Amplification and overexpression of TBX2 in human breast cancer cells.

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Acknowledgements

We thank K. Berns for providing retroviral library DNA; C.J. Sherr for Cdkn2a (ARF)−/− mice; R.A. DePinho for Cdkn2a (INK4a/ARF)−/− mice; L.A. Donehower for Trp53−/− mice; S. Jones for Mdm2−/−;Trp53−/− MEFs; P. Jones for CDKN2A (ARF) promoter-CAT reporter plasmids; M.Oren for cyclin G-Luc plasmid; G. Zambetti for 2A10 (αMdm2) serum; H. Masselink and M. Hijmans for pCMV-E2F1 plasmid; K. Kieboom, C. Bosch, D. Atsma and J. Poodt for technical assistance; A.H. Lund for the 1.7-kb mouse Cdkn2a (ARF) promoter-Luciferase reporter; and R. Agami and J.-W. Voncken for comments on the manuscript. J.J.L.J, P.K. and E.R.M were supported by grants from the Dutch Cancer Society (K.W.F). G.Q.D was supported by grants from the Edward Mallinckrodt, Jr. Foundation and NCI CA76418-01.

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Authors and Affiliations

  1. Division of Molecular Carcinogenesis, The Netherlands Cancer Institute, Amsterdam, The Netherlands

    Jacqueline J.L. Jacobs, Petra Keblusek, Merel Lingbeek & Maarten van Lohuizen

  2. Department of Pathology, The Netherlands Cancer Institute, Amsterdam, The Netherlands

    Els Robanus-Maandag, Petra Kristel, Petra M. Nederlof, Tibor van Welsem & Marc J. van de Vijver

  3. Division of Experimental Therapy, The Netherlands Cancer Institute, Amsterdam, The Netherlands

    Els Robanus-Maandag, Petra Kristel & Marc J. van de Vijver

  4. Whitehead Institute, Cambridge, Massachusetts, USA

    Eugene Y. Koh & George Q. Daley

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Correspondence to Maarten van Lohuizen.

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Jacobs, J., Keblusek, P., Robanus-Maandag, E. et al. Senescence bypass screen identifies TBX2, which represses Cdkn2a (p19ARF) and is amplified in a subset of human breast cancers. Nat Genet 26, 291–299 (2000). https://doi.org/10.1038/81583

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