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Effects of metformin on breast cancer cell proliferation, the AMPK pathway and the cell cycle

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Abstract

Aim

The aim of this study was to compare the effects and mechanisms of action of metformin on estrogen receptor (ER)-positive and ER-negative breast cancer cell lines.

Methods

The anti-proliferative effects of metformin, and of the direct activator of adenosine monophosphate-activated protein kinase (AMPK), A-769662, on MCF-7 (ER-positive) and MDA-MB-231 (ER-negative) breast cancer cell lines were evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, a yellow tetrazole) assays. Fluorescence-activated cell sorting was also used to examine the effect of metformin on the cell cycle. Finally, phosphorylation of the metformin target AMPK, and of its potential downstream targets including acetyl-CoA carboxylase (ACC), p53, p70-S6K and Raptor, was examined using immunoblotting.

Results

Metformin and A-769662 caused significant, concentration-dependent suppression of cell proliferation with G1 cell cycle arrest in both MCF-7 and MDA-MB-231 cells. The proliferation suppression effect was more profound in MCF-7 cells. A concentration-dependent phosphorylation of AMPK was detected following metformin treatment, as was phosphorylation of ACC in both cell lines, but not p53, p70-S6k or Raptor.

Conclusion

Metformin acts as a growth inhibitor in both ER-positive and ER-negative breast cancer cells in vitro, and arrests cells in G1 phase, particularly in the ER-positive MCF-7 cells. The effect is likely to be mediated by AMPK activation, in part by inhibition of fatty acid synthesis via ACC phosphorylation.

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Acknowledgments

All authors acknowledge help and contributions from: AMT group, in particular Ms Karen Murray. DGH group, in particular Dr. Jamie Sampayo, Dr. Sarah Fogarty, Dr. Simon Hawley, Dr. Mhairi Towler, Dr. Fiona Ross and Mr. Kevin Green. Breast Cancer Research Scotland for funding the project.

Conflict of interest

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Academic Unit of Surgical Oncology, Department of Oncology, The Medical School, University of Sheffield, Sheffield, S10 2RX, UK

    S. M. Hadad

  2. Division of Cell Signalling and Immunology, College of Life Sciences, University of Dundee, Dundee, UK

    D. G. Hardie

  3. Clinical Research Centre, Dundee Cancer Centre, University of Dundee, Dundee, UK

    V. Appleyard & A. M. Thompson

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  1. S. M. Hadad
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  2. D. G. Hardie
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  3. V. Appleyard
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Correspondence to S. M. Hadad.

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Hadad, S.M., Hardie, D.G., Appleyard, V. et al. Effects of metformin on breast cancer cell proliferation, the AMPK pathway and the cell cycle. Clin Transl Oncol 16, 746–752 (2014). https://doi.org/10.1007/s12094-013-1144-8

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  • DOI: https://doi.org/10.1007/s12094-013-1144-8

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