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Combined inhibition of glycolysis and AMPK induces synergistic breast cancer cell killing

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Abstract

Targeting glycolysis for cancer treatment has been investigated as a therapeutic method but has not offered a feasible chemotherapeutic strategy. Our aim was to examine whether AMP-activated protein kinase (AMPK), a conditional oncogene, rescues the energetic stress and cytotoxicity induced by 2-deoxyglucose (2-DG), a glycolytic inhibitor, and the related mechanisms. Luciferin/luciferase adenosine triphosphate (ATP) determination, Western analysis, qRT-PCR analyses, MTT growth assay, clonogenic assay, and statistical analysis were performed in this study. 2-DG decreased ATP levels and subsequently activated AMPK, which contribute to intracellular ATP recovery in MCF-7 cells thus exhibiting no apparent cytotoxicity. Compound C, an AMPK inhibitor, further potentiates 2-DG-induced decrease in ATP levels and inhibits their recovery. 2-DG, via AMPK activation, stimulated cAMP response element-binding protein (CREB) phosphorylation and activity and promoted nuclear peroxisome proliferator-activated receptor gamma coactivator-1-beta (PGC-1β) and estrogen-related receptor α (ERRα) protein expression, leading to augmented mitochondrial biogenesis and expression of fatty acid oxidation (FAO) genes including PPARα, MCAD, CPT1C, and ACO. This metabolic adaptation elicited by AMPK counteracts the ATP-depleting and cancer cell-killing effect of 2-DG. However, 2-DG in combination with AMPK antagonists or small interfering RNA caused a dramatic increase in cytotoxicity in MCF-7 but not in MCF-10A cells. Similarly, when combined with inhibition of CREB/PGC-1β/ERRα pathway, 2-DG saliently suppressed mitochondrial biogenesis and the expression of FAO genes, depleted ATP production, and enhanced cytotoxicity in cancer cells. Collectively, the combination of 2-DG and AMPK inhibition synergistically enhanced the cytotoxic potential in breast cancer cells with a relative nontoxicity to normal cells and may offer a promising, safe, and effective breast cancer therapeutic strategy.

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Acknowledgments

This work was supported by the National Institutes of Health (NIH, NCI, NIMHD, NCATS) Grants: U54 CA143931-01, U54MD007598, UL1TR000124 (J. V. Vadgama).

Conflict of interest

All authors declare no conflict of interest.

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Authors and Affiliations

  1. Division of Cancer Research and Training, Charles R. Drew University of Medicine and Science, Los Angeles, CA, 90059, USA

    Yong Wu, Marianna Sarkissyan, Eva Mcghee & Jaydutt V. Vadgama

  2. David Geffen School of Medicine, University of California, Los Angeles, CA, 90095, USA

    Yong Wu & Jaydutt V. Vadgama

  3. Department of Clinical Pharmacy, University of California San Francisco, San Francisco, CA, 94131, USA

    Eva Mcghee

  4. Department of Biochemistry, University of California, Riverside, CA, 92521, USA

    Sangkyu Lee

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  1. Yong Wu
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  5. Jaydutt V. Vadgama
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Correspondence to Jaydutt V. Vadgama.

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Wu, Y., Sarkissyan, M., Mcghee, E. et al. Combined inhibition of glycolysis and AMPK induces synergistic breast cancer cell killing. Breast Cancer Res Treat 151, 529–539 (2015). https://doi.org/10.1007/s10549-015-3386-3

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  • DOI: https://doi.org/10.1007/s10549-015-3386-3

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