Suppression of tumorigenesis by the p53 target PUMA
Abstract
The p53 tumor suppressor regulates diverse antiproliferative processes such that cells acquiring p53 mutations have impaired cell-cycle checkpoints, senescence, apoptosis, and genomic stability. Here, we use stable RNA interference to examine the role of PUMA, a p53 target gene and proapoptotic member of the Bcl2 family, in p53-mediated tumor suppression. PUMA short hairpin RNAs (shRNAs) efficiently suppressed PUMA expression and p53-dependent apoptosis but did not impair nonapoptotic functions of p53. Like p53 shRNAs, PUMA shRNAs promoted oncogenic transformation of primary murine fibroblasts by the E1A/ras oncogene combination and dramatically accelerated myc-induced lymphomagenesis without disrupting p53-dependent cell-cycle arrest. However, the ability of PUMA to execute p53 tumor suppressor functions was variable because, in contrast to p53 shRNAs, PUMA shRNAs were unable to cooperate with oncogenic ras in transformation. These results demonstrate that the p53 effector functions involved in tumor suppression are context dependent and, in some settings, depend heavily on the expression of a single proapoptotic effector. Additionally, they demonstrate the utility of RNA interference for evaluating putative tumor suppressor genes in vivo.
Acknowledgments
We thank L. Bianco for histology; R. Sachidanandam for bioinformatics support; C. Rosenthal, M.-M. Yang, and S. Ray for technical assistance; and R. Dickins and other members of the Lowe laboratory for advice and critical reading of the manuscript. This work was supported by a postdoctoral fellowship from the Helen Hay Whitney Foundation (to M.T.H.), a postdoctoral training grant (to J.T.Z.), National Cancer Institute Project Grants CA13106 (to G.J.H. and S.W.L.) and CA87497 (to S.W.L.), and a generous gift from the Laurie Strauss Leukemia Foundation. D.J.B. is the recipient of a Watson School of Biological Sciences Engelhorn Fellowship, and S.W.L. is an American Association for Cancer Research–National Foundation for Cancer Research Research Professor.
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Copyright © 2004, The National Academy of Sciences.
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Received: April 4, 2004
Published online: June 10, 2004
Published in issue: June 22, 2004
Acknowledgments
We thank L. Bianco for histology; R. Sachidanandam for bioinformatics support; C. Rosenthal, M.-M. Yang, and S. Ray for technical assistance; and R. Dickins and other members of the Lowe laboratory for advice and critical reading of the manuscript. This work was supported by a postdoctoral fellowship from the Helen Hay Whitney Foundation (to M.T.H.), a postdoctoral training grant (to J.T.Z.), National Cancer Institute Project Grants CA13106 (to G.J.H. and S.W.L.) and CA87497 (to S.W.L.), and a generous gift from the Laurie Strauss Leukemia Foundation. D.J.B. is the recipient of a Watson School of Biological Sciences Engelhorn Fellowship, and S.W.L. is an American Association for Cancer Research–National Foundation for Cancer Research Research Professor.
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