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Nitric oxide attenuates resistance to doxorubicin in three-dimensional aggregates of human breast carcinoma cells

  • Preclinical study
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Summary

Compared with monolayer culture, tumour cells cultured as multicellular aggregates (spheroids) exhibit much higher levels of resistance to chemotherapeutic agents, a phenomenon known as multicellular resistance (MCR). Associated with multicellular aggregates is a heterogeneous microenvironment characterised by gradients in oxygen, pH, and nutrients. We previously showed that nitric oxide (NO) signalling plays an important role in the regulation of chemosensitivity in cancer cells cultured as monolayer, and that hypoxia increases resistance to anti-cancer agents largely through a mechanism involving the inhibition of NO signalling. The goal of the present study was to determine whether NO mimetics chemosensitize breast cancer cells in spheroid cultures. Survival of MDA-MB-231 breast carcinoma cells was determined by clonogenic assay following spheroid culture, doxorubicin exposure, and NO mimetic administration. When spheroids were incubated for 24 h with the NO mimetics diethylenetriamine/nitric oxide adduct (DETA/NO) and glyceryl trinitrate (GTN), cell survival after doxorubicin (200 μM) exposure was decreased by 33% (p<0.006) and by up to 47% (p<0.02), respectively. Nitric oxide-mediated signalling involves the generation of the second messenger cyclic guanosine monophosphate (cGMP). Administration of a non-hydrolysable cGMP analogue, 8-Bromo-cGMP, significantly decreased MCR (p<0.04). The effect of NO mimetic exposure on tumour cell chemosensitivity was not due to increased penetration of doxorubicin into spheroids, nor was it associated with an increase in cell proliferation. These results suggest that NO mimetics attenuate MCR to doxorubicin through a mechanism involving cGMP-dependent signalling. Therefore, NO-mimetics may potentially be used as chemosensitizers in cancer therapy.

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Acknowledgements

We thank Ms. Lori Maxwell for her technical support as well as Mr. Matt Gordon for his assistance with the flow cytometric analysis. This work was supported by a grant from the Canadian Institutes of Health Research and a research contract from Cellegy Pharmaceuticals Inc.

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Authors and Affiliations

  1. Department of Anatomy and Cell Biology, Queen’s University, Kingston, ON, Canada

    C.P. Muir & C.H. Graham

  2. Department of Pharmacology and Toxicology, Queen’s University, Kingston, ON, Canada

    M.A. Adams & C.H. Graham

  3. Department of Anatomy and Cell Biology, Queen’s University, Botterell Hall Room 859, K7L 3N6, Kingston, ON, Canada

    C.H. Graham

Authors
  1. C.P. Muir
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  2. M.A. Adams
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  3. C.H. Graham
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Correspondence to C.H. Graham.

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Muir, C., Adams, M. & Graham, C. Nitric oxide attenuates resistance to doxorubicin in three-dimensional aggregates of human breast carcinoma cells. Breast Cancer Res Treat 96, 169–176 (2006). https://doi.org/10.1007/s10549-005-9076-9

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  • DOI: https://doi.org/10.1007/s10549-005-9076-9

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