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Expression and functional analysis of G1 to S regulatory components reveals an important role for CDK2 in cell cycle regulation in human embryonic stem cells

Oncogene volume 28, pages 20–30 (2009)Cite this article

Abstract

One of the characteristic features of human embryonic stem cells (hESCs) is the competence for self-renewal and pluripotency. To date, little is known about cell cycle regulation in these cells and how the cell cycle machinery influences hESCs properties. A common feature of human, murine and primate ESCs is the presence of a short G1 phase, which has been viewed as a time window during which stem cells are exposed to differentiation signals. We used the hESCs differentiation model and comparisons to human embryonic carcinoma (EC) cells to study the key regulators of G1 to S transition in hESCs. Our studies show that hESCs express all G1-specific CYCLINs (D1, D2, D3 and E) and cyclin-dependent kinases (CDK) (CDK2, CDK4 and CDK6) at variable levels. In contrast to murine ESCs, most of the cell cycle regulators in hESCs show cell cycle-dependent expression, thus revealing important differences in the expression of cell cycle regulatory components between these two embryonic cell types. Knockdown of CDK2 using RNA interference resulted in hESCs arrest at G1 phase of the cell cycle and differentiation to extraembryonic lineages. This suggests an important role for CDK2 in cell cycle regulation in hESCs that are likely to bear significant impacts on the maintenance of their pluripotent phenotype.

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Acknowledgements

We are grateful to Mr I Dimmick and Dr R Stewart for help with the flow cytometry analysis, Dr A Hampl for useful discussions, A Khnykina for help with figure preparation, Dr S Pryzborski for providing the EC cells, and Mr G Anyfantis and Mr D Kirk for technical assistance. This study was supported by BBSRC Grant no. BBS/B/14779, MRC Grant G0301182 and Newcastle University.

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Authors and Affiliations

  1. North East Institute for Stem Cell Research, University of Newcastle upon Tyne, International Centre for Life, Newcastle upon Tyne, UK

    I Neganova, X Zhang, S Atkinson & M Lako

  2. Institute of Human Genetics, University of Newcastle upon Tyne, International Centre for Life, Newcastle upon Tyne, UK

    I Neganova, X Zhang, S Atkinson & M Lako

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  1. I Neganova
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  2. X Zhang
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  4. M Lako
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Correspondence to M Lako.

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

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Neganova, I., Zhang, X., Atkinson, S. et al. Expression and functional analysis of G1 to S regulatory components reveals an important role for CDK2 in cell cycle regulation in human embryonic stem cells. Oncogene 28, 20–30 (2009). https://doi.org/10.1038/onc.2008.358

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