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Differentiation

Effects of 5-aza-2′-deoxycytidine on proliferation, differentiation and p15/INK4b regulation of human hematopoietic progenitor cells

Leukemia volume 20, pages 115–121 (2006)Cite this article

Abstract

The demethylating agents 5-azacytidine and 5-aza-2′-deoxycytidine (DAC) have been shown to induce differentiation and inhibit growth of leukemic myeloid cells at low concentrations. However, the effect of DAC in changing the differentiation and proliferation behavior of normal human myeloid progenitors has rarely been investigated. Therefore, we established an in vitro model of normal hematopoietic differentiation, using CD34+ cells from mobilized peripheral blood, to study proliferation and colony formation, expression of several myeloid maturation markers and of the inhibitor of cyclin-dependent kinases p15/INK4b. Upon DAC treatment, cell growth was significantly decreased in a dose-dependent manner, without an increase in cytotoxicity. DAC treatment also resulted in a substantial increase of lysozyme-positive cells, which could be enhanced by G-CSF, a modest increase of myeloperoxidase+ and CD15+ cells, as well as an increase of colony-forming cells (CFU-GM) compared to control cells. p15/INK4b protein expression was strongly upregulated upon myeloid maturation, and additional DAC treatment did not change p15 expression or the methylation status of the p15 promoter at the noncytotoxic concentrations used. Taken together, these data indicate a role of DAC in changing myeloid progenitor cell expansion and differentiation. This model appears suitable also for global analyses of multiple differentially methylated genes.

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Acknowledgements

We thank Eva Samek for excellent technical support and Dr Jonathan Dodge, Boston, for valuable suggestions during the performance of experiment. We are grateful to Professor Dr Fusenig (German Cancer Center) for providing the HaCat cell line. We greatly acknowledge Drs Florian Otto, Ralph Wäsch, Tobias Berg and Michael Stock for valuable suggestions and critical reading of the manuscript, appreciate Professor Dr Roland Mertelsmann's continuous support, and Professor Dr Thomas Licht's helpful discussion. We specially thank Dr Diane Church (Dartmouth Medical School) for expert assistance. This work was supported by a grant from Deutsche Forschungsgemeinschaft (Lu 429/5-1).

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Authors and Affiliations

  1. Department of Hematology/Oncology, University of Freiburg Medical Center, Freiburg, Germany

    Y Guo, M Engelhardt, D Wider, M Abdelkarim & M Lübbert

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  1. Y Guo
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  2. M Engelhardt
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Correspondence to M Lübbert.

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Guo, Y., Engelhardt, M., Wider, D. et al. Effects of 5-aza-2′-deoxycytidine on proliferation, differentiation and p15/INK4b regulation of human hematopoietic progenitor cells. Leukemia 20, 115–121 (2006). https://doi.org/10.1038/sj.leu.2404019

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