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E2F-1 transcriptional activity is a critical determinant of Mdm2 antagonist-induced apoptosis in human tumor cell lines

Oncogene volume 27, pages 5303–5314 (2008)Cite this article

Abstract

Nutlin-3 is a selective inhibitor of the p53-Mdm2 interaction, and inhibits growth in most tumor cells with wild-type p53. However, it only induces apoptosis in subsets of tumor cells. We report that the apoptotic response induced by Nutlin-3 correlates with its antitumor effects in xenograft models in athymic mice. We have investigated signals that sensitize cells to undergo apoptosis induced by Nutlin-3. We demonstrate that adenovirus E1A increases Nutlin-3-induced apoptosis through pRb inhibition in mouse embryonic fibroblast cells in a p53-dependent manner. Consistent with this, pRb depletion by siRNA transfection with Nutlin-3 synergistically increases apoptosis in HCT116 human colon cancer cells, which are insensitive to induction of apoptosis by Nutlin-3 alone. As pRb is a key negative regulator of E2F, we asked whether E2F transcriptional activity determines the apoptotic response of cancer cells to Nutlin-3. We demonstrate that transcriptional activity of E2F correlates with the apoptotic response to Nutlin-3 in various tumor cells and depletion of E2F-1 suppresses Nutlin-3-induced apoptosis in cells possessing high transcriptional activity of E2F, including retinoblastoma cells harboring mutated Rb with wild-type p53. Furthermore, we report that expression of the p53 homologue p73, a target of E2F-1, is markedly increased by Nutlin-3 in Rb-mutated tumor cells harboring wild-type p53. Depletion of p73 by siRNA transfection suppresses Nutlin-3-induced apoptosis in these cells. Taken together, our results demonstrate that E2F-1 transcriptional activity is a critical determinant of Mdm2 antagonist-induced apoptosis and p73 is important for E2F-1-mediated apoptosis induced by Nutlin-3, especially in tumor cells with mutated Rb. Furthermore, our results suggest that tumor cells, including Rb mutated cells, which harbor wild-type p53 and high E2F transcriptional activity, could be a good target for Mdm2 antagonist therapy.

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Acknowledgements

We thank Dr Andy Finch (UCSF), Dr Martin McMahon (UCSF) and Ms Amy Young in the McCormick laboratory for sharing reagents; Daiichi Sankyo members: Ms Megumi Minami, Dr Takahiko Seki and Dr Takeshi Jimbo for technical supports, and Dr Hiroyuki Fujiwara and Ms Etsuko Seno for help in plasmid constructions. We acknowledge Dr Gerard Evan (UCSF), Dr Mike Fried (UCSF) and Dr Hideyuki Saya (Keio University) for reading the manuscript carefully and for providing many insightful comments. We also thank the UCSF Comprehensive Cancer Center Genome core facilities for real-time Q-PCR analysis.

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  1. Helen Diller Family Comprehensive Cancer Center and Cancer Research Institute, University of California, San Francisco, CA, USA

    M Kitagawa, S H Lee & F McCormick

  2. Kasai R&D Center, Daiichi Sankyo Co. Ltd, Tokyo, Japan

    M Kitagawa, M Aonuma & S Fukutake

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  1. M Kitagawa
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  2. M Aonuma
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  5. F McCormick
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Correspondence to F McCormick.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

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Kitagawa, M., Aonuma, M., Lee, S. et al. E2F-1 transcriptional activity is a critical determinant of Mdm2 antagonist-induced apoptosis in human tumor cell lines. Oncogene 27, 5303–5314 (2008). https://doi.org/10.1038/onc.2008.164

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