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Demethylating agent 5-aza-2-deoxycytidine enhances susceptibility of breast cancer cells to anticancer agents

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Abstract

DNA methylation plays an important role in regulation of gene expression and is increasingly being recognized as a determinant of chemosensitivity of human cancers. With the aim of improving the chemotherapeutic efficacy of breast carcinoma, the effect of DNA methyltransferase inhibitor, 5-Aza-2′-deoxycytidine (5-aza-CdR), on the chemosensitivity of anticancer drugs was investigated. The cytotoxicity of paclitaxel (PTX), adriamycin (ADR), and 5-fluorouracil (5-FU) was analyzed against human breast cancer cell lines, MDA MB 231 and MCF 7 cell lines using the MTT assay, and the synergy of 5-aza-CdR and these agents was determined by Drewinko’s fraction method. The effects of each single agent or the combined treatment on cell cycle arrest were analyzed by flow cytometric analysis. We also investigated the effect of each single agent or the combined treatment of anticancer drugs with 5-aza-CdR on the methylation status of the selected genes by methylation specific PCR. In MDA MB 231 cells, a synergistic antiproliferative effect was observed with a combination of 10 μM 5-aza-CdR and these three anticancer drugs, while in MCF 7 cells, a semiadditive effect was observed. Treatment with 5-aza-CdR and anticancer drug resulted in partial demethylation of a panel of genes including RARβ2, Slit2, GSTP1, and MGMT. Based on these findings, we propose that 5-aza-CdR enhances the chemosensitivity of anticancer drugs in breast cancer cells and may be a promising approach for increasing the chemotherapeutic potential of these anticancer agents for more effective management of breast carcinomas.

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Acknowledgments

S.M. is thankful to UGC for providing him SRF Fellowship.

Conflicts of Interest

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Authors and Affiliations

  1. Department of Biochemistry, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, 110029, India

    Sameer Mirza, Gayatri Sharma & Ranju Ralhan

  2. Dev Sanskriti Vishwa Vidhyalaya, Haridwar, India

    Gayatri Sharma & Pranav Pandya

  3. Department of Otolaryngology-Head and Neck Surgery, Mount Sinai Hospital, University of Toronto, Toronto, ON, Canada

    Ranju Ralhan

  4. Joseph and Mildred Sonshine Family Centre for Head & Neck Diseases, Mount Sinai Hospital, Joseph & Wolf Lebovic Health Complex, 600 University Avenue, Room 6-500, Toronto, ON, M5G 1X5, Canada

    Ranju Ralhan

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  1. Sameer Mirza
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  2. Gayatri Sharma
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  4. Ranju Ralhan
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Corresponding author

Correspondence to Ranju Ralhan.

Additional information

Sameer Mirza and Gayatri Sharma have contributed equally to this manuscript.

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Mirza, S., Sharma, G., Pandya, P. et al. Demethylating agent 5-aza-2-deoxycytidine enhances susceptibility of breast cancer cells to anticancer agents. Mol Cell Biochem 342, 101–109 (2010). https://doi.org/10.1007/s11010-010-0473-y

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  • DOI: https://doi.org/10.1007/s11010-010-0473-y

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