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Transcriptional upregulation of p57 (Kip2) by the cyclin-dependent kinase inhibitor BMS-387032 is E2F dependent and serves as a negative feedback loop limiting cytotoxicity

Oncogene volume 26, pages 3532–3540 (2007)Cite this article

Abstract

In spite of the fact that cyclin-dependent kinase (cdk) inhibiting drugs are potent transcriptional repressors, we discover that p57 (Kip2, CDKN1C) transcription is significantly upregulated by three small molecule cdk inhibitors, including BMS-387032. Treatment of MDA-MB-231 breast cancer cells with BMS-387032 led to a stabilization of the E2F1 protein that was accompanied by significant increases in the p57 mRNA and protein. This increase did not occur in an E2F1-deficient cell line. An E2F1-estrogen receptor fusion protein activated the endogenous p57 promoter in response to hydroxytamoxifen treatment in the presence of cycloheximide. Luciferase constructs driven by the p57 promoter verified that upregulation of p57 mRNA by BMS-387032 is transcriptional and dependent on E2F-binding sites in the promoter. Expression of exogenous p57 significantly decreased the fraction of cells in S phase. Furthermore, p57-deficient MDA-MB-231 cell lines were significantly more sensitive to BMS-387032-induced apoptosis than controls. The results presented in this manuscript demonstrate that small molecule cdk inhibitors transcriptionally activate p57 dependent upon E2F1 and that this activation in turn serves to limit E2F1's death-inducing activity.

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Acknowledgements

We thank Dr Jack Pledger, Scott Freeman and Jose Rodriguez for scientific input. We also acknowledge Gabriela Wright and Monica Dickinson for performing many experiments. In addition, we thank numerous colleagues for gifts of essential reagents and materials (see Methods and materials). This work was supported by funds from the National Cancer Institute (CA90489-01, WDC) and by the Molecular Biology, Flow Cytometry, Analytical Microscopy, and the Molecular Imaging Core Facilities of the Moffitt Research Institute.

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  1. Molecular Oncology Program, H. Lee Moffitt Cancer Center and Research Institute, University of South Florida, College of Medicine, Tampa, FL, USA

    Y Ma & W D Cress

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Correspondence to W D Cress.

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

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Ma, Y., Cress, W. Transcriptional upregulation of p57 (Kip2) by the cyclin-dependent kinase inhibitor BMS-387032 is E2F dependent and serves as a negative feedback loop limiting cytotoxicity. Oncogene 26, 3532–3540 (2007). https://doi.org/10.1038/sj.onc.1210143

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