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.
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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Molecular Oncology Program, H Lee Moffitt Cancer Center and Research Institute, University of South Florida, College of Medicine, Tampa, FL 33612-9497, USA.
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D001665:Binding Sites; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D050761:Cyclin-Dependent Kinase Inhibitor p57 / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors*; D050684:E2F Transcription Factors / Q000187:drug effects / Q000378:metabolism*; D050687:E2F1 Transcription Factor / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D025461:Feedback, Physiological; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D010080:Oxazoles / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D013844:Thiazoles / Q000494:pharmacology*; D014158:Transcription, Genetic; D015854:Up-Regulation
C493964:CDKN1C protein, human; D050761:Cyclin-Dependent Kinase Inhibitor p57; D050684:E2F Transcription Factors; D050687:E2F1 Transcription Factor; C493753:E2F1 protein, human; D004791:Enzyme Inhibitors; C484864:N-(5-(((5-(1,1-dimethylethyl)-2-oxazolyl)methyl)thio)-2-thiazolyl)-4-piperidinecarboxamide; D010080:Oxazoles; D013844:Thiazoles; D018844:Cyclin-Dependent Kinases