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Mode of interaction between butyroyloxymethyl-diethyl phosphate (AN-7) and doxorubicin in MCF-7 and resistant MCF-7/Dx cell lines

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Abstract

Purpose

To investigate the anticancer activity and mode of action of butyroyloxymethyl-diethyl phosphate (AN-7), a prodrug of butyric acid and formaldehyde, as a single agent and in combination with doxorubicin in human carcinoma MCF-7 and the multidrug resistant MCF-7 Dx cell lines.

Methods

The anti-cancer activity of AN-7 as a single agent or in combination with doxorubicin was measured by the Hoechst cell viability and colony forming assays as well as by FACS analyses of cells stained with propidium iodide and annexin V-FITC. Modulations of protein expression and acetylation were measured by Western blot analyses. The number of doxorubicin–DNA adducts formed was evaluated using 14C-labeled doxorubicin.

Results

The AN-7 and homologous prodrugs exhibited similar growth inhibition effects against drug resistant and sensitive cells, and elicited their anticancer effect partially by inhibition of HDAC. The AN-7 transiently augmented histone acetylation and increase of p21 expression. Synergy between AN-7 and doxorubicin was demonstrated in the sensitive and the resistant cell lines by viability and colony formation assays and was further confirmed by FACS analysis showing an increase in cell mortality. The number of doxorubicin–DNA adducts in total genomic DNA isolated from cells treated with 14C-labeled doxorubicin and AN-7 increased substantially compared to treatment with doxorubicin only. Treatment with AN-7 or doxorubicin increased p53 acetylation that was further potentiated by their combination.

Conclusion

The AN-7 combined with doxorubicin overcame drug resistance; at least in part by the intracellularly releasable formaldehyde that augmented formation of doxorubicin–DNA adducts and butyric acid that induced histone and p53 acetylation. Since the use of doxorubicin is limited by toxicity, the combination could offer an effective treatment modality with lower toxicity for breast cancer.

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Author notes

  1. Michal Entin-Meer

    Present address: Department of Radiation Oncology and Comprehensive Cancer Center, University of California, San Francisco, CA, 94143, USA

Authors and Affiliations

  1. Chemistry Department, Bar-Ilan University, Ramat Gan, 52900, Israel

    Dikla Engel, Abraham Nudelman & Inesa Levovich

  2. Felsenstein Medical Research Center, Sackler School of Medicine, Tel Aviv University Beilinson Campus, Petach Tikva, 49100, Israel

    Dikla Engel, Inesa Levovich, Tal Gruss-Fischer, Michal Entin-Meer & Ada Rephaeli

  3. Department of Biochemistry, La Trobe University, Melbourne, VIC, 3086, Australia

    Don R. Phillips & Suzanne M. Cutts

  4. Head of Pharmacology and Experimental Oncology Laboratory, Felsenstein Medical Research Center, Tel Aviv University Beilinson Campus, Petach Tikva, 49100, Israel

    Ada Rephaeli

Authors
  1. Dikla Engel
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  2. Abraham Nudelman
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  3. Inesa Levovich
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  4. Tal Gruss-Fischer
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  5. Michal Entin-Meer
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  6. Don R. Phillips
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  8. Ada Rephaeli
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Corresponding author

Correspondence to Ada Rephaeli.

Additional information

This work is supported by a grant 542/00-3 from Israel Science Foundation (A.R. and A.N.); a project grant from the Israel Cancer Research Fund, Israel Cancer Association; The Marcus Center for Pharmaceutical and Medicinal Chemistry at Bar Ilan University (A.N.), Australian Research Council (S.M.C. and D.R.P.). This work was performed by D. Engel as part of the requirements for Ph.D. degree in Bar Ilan University

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Engel, D., Nudelman, A., Levovich, I. et al. Mode of interaction between butyroyloxymethyl-diethyl phosphate (AN-7) and doxorubicin in MCF-7 and resistant MCF-7/Dx cell lines. J Cancer Res Clin Oncol 132, 673–683 (2006). https://doi.org/10.1007/s00432-006-0116-6

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  • DOI: https://doi.org/10.1007/s00432-006-0116-6

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