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Plasminogen activator inhibitor-1 is a critical downstream target of p53 in the induction of replicative senescence

Nature Cell Biology volume 8, pages 877–884 (2006)Cite this article

Abstract

p53 limits the proliferation of primary diploid fibroblasts by inducing a state of growth arrest named replicative senescence — a process which protects against oncogenic transformation and requires integrity of the p53 tumour suppressor pathway1,2,3. However, little is known about the downstream target genes of p53 in this growth-limiting response. Here, we report that suppression of the p53 target gene encoding plasminogen activator inhibitor-1 (PAI-1) by RNA interference (RNAi) leads to escape from replicative senescence both in primary mouse embryo fibroblasts and primary human BJ fibroblasts. PAI-1 knockdown results in sustained activation of the PI(3)K–PKB–GSK3β pathway and nuclear retention of cyclin D1, consistent with a role for PAI-1 in regulating growth factor signalling. In agreement with this, we find that the PI(3)K–PKB–GSK3β–cyclin D1 pathway is also causally involved in cellular senescence. Conversely, ectopic expression of PAI-1 in proliferating p53-deficient murine or human fibroblasts induces a phenotype displaying all the hallmarks of replicative senescence. Our data indicate that PAI-1 is not merely a marker of senescence, but is both necessary and sufficient for the induction of replicative senescence downstream of p53.

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Figure 1: PAI-1 loss induces senescence-bypass in primary mouse fibroblasts.
Figure 2: MEFs reduce PKB activation and exclude cyclin D1 from the nucleus during replicative senescence.
Figure 3: Sustained PI(3)K–PKB signalling or nuclear retention of cyclin D1 induces bypass of senescence.
Figure 4: PAI-1 expression is sufficient for the induction of replicative senescence.
Figure 5: PAI-1 is necessary and sufficient for senescence in human BJ fibroblasts.

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Acknowledgements

We would like to thank A. Visser for technical assistance, K. Berns, M. Hijmans, A. Dirac, T. Brummelkamp, R. Agami, R. van der Kammen, J. Collard and F. Scheeren for retroviral constructs, B. Weigelt for help with QRT–PCR, F. Foijer for retinoblastoma family deficient MEFs, L. Oomen and L. Brocks for help with microscopy, and R. Beijersbergen and D. Peeper for helpful discussions. This work was supported by a grant from the Dutch Cancer Society to R.B. and a grant from the National Institutes of Health (NIH; GM57242) to P.H.

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Authors and Affiliations

  1. Division of Molecular Carcinogenesis and Center for Biomedical Genetics, The Netherlands Cancer Institute, Amsterdam, 1211066 CX, Plesmanlaan, The Netherlands

    Roderik M. Kortlever & René Bernards

  2. Center for Cell Biology & Cancer Research, Albany Medical College, MC-165, 47 New Scotland Avenue, Albany, NY 12208, USA

    Paul J. Higgins

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R.K. and R.B. conceived and designed the experiments. R.K. performed the experiments. P.H. contributed materials. R.K. and R.B. analysed the data. R.K. and R.B. wrote the paper.

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Correspondence to René Bernards.

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Kortlever, R., Higgins, P. & Bernards, R. Plasminogen activator inhibitor-1 is a critical downstream target of p53 in the induction of replicative senescence. Nat Cell Biol 8, 877–884 (2006). https://doi.org/10.1038/ncb1448

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