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  • Original Paper
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De novo DNA methyltransferases Dnmt3a and Dnmt3b primarily mediate the cytotoxic effect of 5-aza-2′-deoxycytidine

Oncogene volume 24, pages 3091–3099 (2005)Cite this article

Abstract

The deoxycytidine analog 5-aza-2′-deoxycitidine (5-aza-dC) is a potent chemotherapeutic agent effective against selective types of cancer. The molecular mechanism by which 5-aza-dC induces cancer cell death, however, is not fully understood. It has been accepted that the mechanism of toxicity is due to the covalent binding between the DNA methyltransferase (Dnmt) and 5-aza-dC-substituted DNA. In order to define which member of the Dnmt family plays a dominant role in the cytotoxicity, we examined the effect of 5-aza-dC on cell growth and apoptosis in various Dnmt null mutant embryonic stem (ES) cells. Of interest, Dnmt3a–Dnmt3b double null ES cells were highly resistant to 5-aza-dC when compared to wild type, Dnmt3a null, Dnmt3b null, or Dnmt1 null ES cells. The cellular sensitivity to 5-aza-dC correlated well with the expression status of Dnmt3 in both undifferentiated and differentiated ES cells. When exogenous Dnmt3a or Dnmt3b was expressed in double null ES cells, the sensitivity to 5-aza-dC was partially restored. These results suggest that the cytotoxic effect of 5-aza-dC may be mediated primarily through Dnmt3a and Dnmt3b de novo DNA methyltransferases. Further, the ability to form Dnmt-DNA adducts was similar in Dnmt1 and Dnmt3, and the expression level of Dnmt3 was not higher than that of Dnmt1 in ES cells. Therefore, Dnmt3-DNA adducts may be more effective for inducing apoptosis than Dnmt1-DNA adducts. These results imply a therapeutic potential of 5-aza-dC to cancers expressing Dnmt3.

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Acknowledgements

We thank Dr En Li for providing Dnmt null ES cells, Dr Shoji Tajima for anti-Dnmt1 antibody, and Dr Keith D Robertson and Dr Thomas C Rowe for critical reading of the manuscript.

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Authors and Affiliations

  1. Department of Pathology, University of Florida College of Medicine, PO Box 100275, 1600 SW Archer Rd, Gainesville, FL, 32610, USA

    Masahiro Oka, Amy M Meacham, Takashi Hamazaki, Nemanja Rodić & Naohiro Terada

  2. Department of Molecular Genetics and Microbiology, Powell Gene Therapy Center and McKnight Brain Institute, University of Florida, Gainesville, FL, 32610, USA

    Lung-Ji Chang

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  1. Masahiro Oka
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Correspondence to Naohiro Terada.

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Oka, M., Meacham, A., Hamazaki, T. et al. De novo DNA methyltransferases Dnmt3a and Dnmt3b primarily mediate the cytotoxic effect of 5-aza-2′-deoxycytidine. Oncogene 24, 3091–3099 (2005). https://doi.org/10.1038/sj.onc.1208540

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