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Enhancement of chemotherapeutic efficacy in hypermethylator breast cancer cells through targeted and pharmacologic inhibition of DNMT3b

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Abstract

A subset of primary breast cancers and breast cancer cell lines express a hypermethylation defect (characterized by DNMT hyperactivity and DNMT3b overexpression) which contributes to chemotherapy resistance and provides a target for development of new treatment strategies. The objective of the current study was to determine if targeting the epigenome enhances the sensitivity of breast cancer cells to cytotoxic chemotherapy. Hypermethylator breast cancer cell lines (MDA-MB-453, BT549, and Hs578T) were treated with 250 or 500 nM 5-aza-2′-deoxycytidine (5-aza) and/or were subjected to RNAi-mediated DNMT3b knockdown (KD), and then tested for sensitivity to doxorubicin hydrochloride (DOX), paclitaxel (PAX), and 5-fluorouracil (5-FU). In MDA-MB-453 cells, DNMT3b KD reduces the IC50 for DOX from 0.086 to 0.048 μM (44% reduction), for PAX from 0.497 to 0.376 nM (24%), and for 5-FU from 0.817 to 0.145 mM (82%). Treatment with 250 nM 5-aza for 7 days did not increase the efficacy of DOX, PAX, or 5-FU, but 7-day treatment with 500 nM 5-aza sensitized cells, reducing the IC50 for DOX to 0.035 μM (60%), PAX to 0.311 nM (37%), and 5-FU to 0.065 mM (92%). 5-aza treatment of DNMT3b KD cells reduced the IC50 for DOX to 0.036 μM (59%), for PAX to 0.313 nM (37%) and for 5-FU to 0.067 (92%). Similar trends of enhancement of cell kill were seen in BT549 (13–60%) and Hs578T (29–70%) cells after RNAi-mediated DNMT3b KD and/or treatment with 5-aza. The effectiveness of DOX, PAX, and 5-FU is enhanced through targeted and/or pharmacological inhibition of DNMT3b, strongly suggesting that combined epigenetic and cytotoxic treatment will improve the efficacy of breast cancer chemotherapy.

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Acknowledgments

This study was supported by a University Cancer Research Fund grant from the UNC Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine. AGR is supported by a Postdoctoral Fellowship grant from the American Cancer Society (PF-08-166-01-GMC).

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The authors declare that there are no conflicts of interest regarding the contents of this article.

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Authors and Affiliations

  1. Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, 515 Brinkhous Bullitt Building, CB #7525, Chapel Hill, NC, 27599, USA

    Rupninder Sandhu & William B. Coleman

  2. Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, NC, USA

    Ashley G. Rivenbark

  3. UNC Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Chapel Hill, NC, USA

    Rupninder Sandhu, Ashley G. Rivenbark & William B. Coleman

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  1. Rupninder Sandhu
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  2. Ashley G. Rivenbark
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Correspondence to William B. Coleman.

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Sandhu, R., Rivenbark, A.G. & Coleman, W.B. Enhancement of chemotherapeutic efficacy in hypermethylator breast cancer cells through targeted and pharmacologic inhibition of DNMT3b. Breast Cancer Res Treat 131, 385–399 (2012). https://doi.org/10.1007/s10549-011-1409-2

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