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Cyclin-dependent kinase 1 expression is inhibited by p16INK4a at the post-transcriptional level through the microRNA pathway

Oncogene volume 30, pages 1880–1891 (2011)Cite this article

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Abstract

The p16INK4a protein regulates cell cycle progression mainly by inhibiting the activity of G1-phase cyclin-dependent kinases (CDKs) 4 and 6, the subsequent retinoblastoma protein (pRb) phosphorylation and E2F transcription factor release. The p16INK4a protein can also repress the activity of other transcription factors, such as c-myc, nuclear factor-kappaB and c-Jun/AP1. Here, we report that, in two p16−/−, pRbWT and p53WT cell lines (MCF7 and U87), p16INK4a overexpression induces a dramatic decrease in CDK1 protein expression. In response to p16INK4a, the decreased rate of CDK1 protein synthesis, its unchanged protein half-life, unreduced CDK1 mRNA steady-state levels and mRNA half-life allow us to hypothesize that p16INK4a could regulate CDK1 expression at the post-transcriptional level. This CDK1 downregulation is mediated by the 3′-untranslated region (3′UTR) of CDK1 mRNA as shown by translational inhibition in luciferase assays and is associated with a modified expression balance of microRNAs (miRNAs) that potentially regulate CDK1, analyzed by TaqMan Human microRNA Array. The p16INK4a-induced expression of two miRNAs (miR-410 and miR-650 chosen as an example) in MCF7 cells is confirmed by individual reverse transcription–qPCR. Furthermore, we show the interaction of miR-410 or miR-650 with CDK1-3′UTR by luciferase assays. Endogenous CDK1 expression decreases upon both miRNA overexpression and increases with their simultaneous inhibition. The induction of miR-410, but not miR-650 could be related to the pRb/E2F pathway. These results demonstrate the post-transcriptional inhibition of CDK1 by p16INK4a. We suggest that p16INK4a may regulate gene expression by modifying the functional equilibrium of transcription factors and consequently the expression balance of miRNAs.

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Acknowledgements

This work was supported by the Ligue contre le Cancer of Saône-et-Loire and Rhône, by the Fondation pour la Recherche Médicale and by the Cancéropole Lyon-Auvergne-Rhône-Alpes.

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Authors and Affiliations

  1. Université Claude Bernard Lyon 1, CNRS UMR 5239 ENS—HCL, Faculté de Médecine Lyon Sud, Oullins, France

    W W Chien, C Domenech, R Catallo, T Kaddar, J-P Magaud, G Salles & M Ffrench

  2. Laboratoire d’Hématologie, CHLS, Hospices Civils de Lyon, 165 Chemin du Grand Revoyet, Pierre-Bénite, France

    J-P Magaud & M Ffrench

  3. Service d’Hématologie, CHLS, Hospices Civils de Lyon, Pierre-Bénite, France

    G Salles

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Correspondence to M Ffrench.

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Chien, W., Domenech, C., Catallo, R. et al. Cyclin-dependent kinase 1 expression is inhibited by p16INK4a at the post-transcriptional level through the microRNA pathway. Oncogene 30, 1880–1891 (2011). https://doi.org/10.1038/onc.2010.570

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