Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Differential effects of AMPK agonists on cell growth and metabolism

Abstract

As a sensor of cellular energy status, the AMP-activated protein kinase (AMPK) is believed to act in opposition to the metabolic phenotypes favored by proliferating tumor cells. Consequently, compounds known to activate AMPK have been proposed as cancer therapeutics. However, the extent to which the anti-neoplastic properties of these agonists are mediated by AMPK is unclear. Here we examined the AMPK dependence of six commonly used AMPK agonists (metformin, phenformin, 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), 2-deoxy-D-glucose (2DG), salicylate and A-769662) and their influence on cellular processes often deregulated in tumor cells. We demonstrate that the majority of these agonists display AMPK-independent effects on cell proliferation and metabolism with only the synthetic activator, A-769662, exerting AMPK-dependent effects on these processes. We find that A-769662 promotes an AMPK-dependent increase in mitochondrial spare respiratory capacity. Finally, contrary to the view of AMPK activity being tumor suppressive, we find that A-769662 confers a selective proliferative advantage to tumor cells growing under nutrient deprivation. Our results indicate that many of the antigrowth properties of these agonists cannot be attributed to AMPK activity in cells, and thus any observed effects using these agonists should be confirmed using AMPK-deficient cells. Ultimately, our data urge caution not only regarding the type of AMPK agonist proposed for cancer treatment but also the context in which they are used.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

References

  1. Suter M, Riek U, Tuerk R, Schlattner U, Wallimann T, Neumann D . Dissecting the role of 5′-AMP for allosteric stimulation, activation, and deactivation of AMP-activated protein kinase. J Biol Chem 2006; 281: 32207–32216.

    Article  CAS  PubMed  Google Scholar 

  2. Hardie DG . AMP-activated/SNF1 protein kinases: conserved guardians of cellular energy. Nat Rev Mol Cell Biol 2007; 8: 774–785.

    Article  CAS  PubMed  Google Scholar 

  3. Carling D . The AMP-activated protein kinase cascade—a unifying system for energy control. Trends Biochem Sci 2004; 29: 18–24.

    Article  CAS  PubMed  Google Scholar 

  4. Hardie DG . AMP-activated protein kinase: a cellular energy sensor with a key role in metabolic disorders and in cancer. Biochem Soc Trans 2011; 39: 1–13.

    Article  CAS  PubMed  Google Scholar 

  5. Davies SP, Sim AT, Hardie DG . Location and function of three sites phosphorylated on rat acetyl-CoA carboxylase by the AMP-activated protein kinase. Eur J Biochem 1990; 187: 183–190.

    Article  CAS  PubMed  Google Scholar 

  6. Gwinn DM, Shackelford DB, Egan DF, Mihaylova MM, Mery A, Vasquez DS et al. AMPK phosphorylation of raptor mediates a metabolic checkpoint. Mol Cell 2008; 30: 214–226.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Inoki K, Zhu T, Guan KL . TSC2 mediates cellular energy response to control cell growth and survival. Cell 2003; 115: 577–590.

    Article  CAS  PubMed  Google Scholar 

  8. Imamura K, Ogura T, Kishimoto A, Kaminishi M, Esumi H . Cell cycle regulation via p53 phosphorylation by a 5'-AMP activated protein kinase activator, 5-aminoimidazole- 4-carboxamide-1-beta-D-ribofuranoside, in a human hepatocellular carcinoma cell line. Biochem Biophys Res Commun 2001; 287: 562–567.

    Article  CAS  PubMed  Google Scholar 

  9. Jones RG, Plas DR, Kubek S, Buzzai M, Mu J, Xu Y et al. AMP-activated protein kinase induces a p53-dependent metabolic checkpoint. Mol Cell 2005; 18: 283–293.

    Article  CAS  PubMed  Google Scholar 

  10. Shackelford DB, Shaw RJ . The LKB1-AMPK pathway: metabolism and growth control in tumour suppression. Nat Rev Cancer 2009; 9: 563–575.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Hawley SA, Boudeau J, Reid JL, Mustard KJ, Udd L, Makela TP et al. Complexes between the LKB1 tumor suppressor, STRADalpha/beta and MO25alpha/beta are upstream kinases in the AMP-activated protein kinase cascade. J Biol 2003; 2: 28.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Alessi DR, Sakamoto K, Bayascas JR . LKB1-dependent signaling pathways. Annu Rev Biochem 2006; 75: 137–163.

    Article  CAS  PubMed  Google Scholar 

  13. Giardiello FM, Welsh SB, Hamilton SR, Offerhaus GJ, Gittelsohn AM, Booker SV et al. Increased risk of cancer in the Peutz-Jeghers syndrome. New Engl J Med 1987; 316: 1511–1514.

    Article  CAS  PubMed  Google Scholar 

  14. Hearle N, Schumacher V, Menko FH, Olschwang S, Boardman LA, Gille JJ et al. Frequency and spectrum of cancers in the Peutz-Jeghers syndrome. Clin Cancer Res 2006; 12: 3209–3215.

    Article  CAS  PubMed  Google Scholar 

  15. Sanchez-Cespedes M . A role for LKB1 gene in human cancer beyond the Peutz-Jeghers syndrome. Oncogene 2007; 26: 7825–7832.

    Article  CAS  PubMed  Google Scholar 

  16. Dupuy F, Griss T, Blagih J, Bridon G, Avizonis D, Ling C et al. LKB1 is a central regulator of tumor initiation and pro-growth metabolism in ErbB2-mediated breast cancer. Cancer Metab 2013; 1: 18.

    Article  PubMed  PubMed Central  Google Scholar 

  17. Shackelford DB, Vasquez DS, Corbeil J, Wu S, Leblanc M, Wu CL et al. mTOR and HIF-1alpha-mediated tumor metabolism in an LKB1 mouse model of Peutz-Jeghers syndrome. Proc Natl Acad Sci USA 2009; 106: 11137–11142.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Faubert B, Boily G, Izreig S, Griss T, Samborska B, Dong Z et al. AMPK is a negative regulator of the Warburg effect and suppresses tumor growth in vivo. Cell Metab 2013; 17: 113–124.

    Article  CAS  PubMed  Google Scholar 

  19. Kim I, He YY . Targeting the AMP-activated protein kinase for cancer prevention and therapy. Front Oncol 2013; 3: 175.

    PubMed  PubMed Central  Google Scholar 

  20. Owen MR, Doran E, Halestrap AP . Evidence that metformin exerts its anti-diabetic effects through inhibition of complex 1 of the mitochondrial respiratory chain. Biochem J 2000; 348 (Pt 3): 607–614.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. El-Mir MY, Nogueira V, Fontaine E, Averet N, Rigoulet M, Leverve X . Dimethylbiguanide inhibits cell respiration via an indirect effect targeted on the respiratory chain complex I. J Biol Chem 2000; 275: 223–228.

    Article  CAS  PubMed  Google Scholar 

  22. Evans JM, Donnelly LA, Emslie-Smith AM, Alessi DR, Morris AD . Metformin and reduced risk of cancer in diabetic patients. BMJ 2005; 330: 1304–1305.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Decensi A, Puntoni M, Goodwin P, Cazzaniga M, Gennari A, Bonanni B et al. Metformin and cancer risk in diabetic patients: a systematic review and meta-analysis. Cancer Prev Res (Phila) 2010; 3: 1451–1461.

    Article  CAS  Google Scholar 

  24. Buzzai M, Jones RG, Amaravadi RK, Lum JJ, DeBerardinis RJ, Zhao F et al. Systemic treatment with the antidiabetic drug metformin selectively impairs p53-deficient tumor cell growth. Cancer Res 2007; 67: 6745–6752.

    Article  CAS  PubMed  Google Scholar 

  25. Wu N, Gu C, Gu H, Hu H, Han Y, Li Q . Metformin induces apoptosis of lung cancer cells through activating JNK/p38 MAPK pathway and GADD153. Neoplasma 2011; 58: 482–490.

    Article  CAS  PubMed  Google Scholar 

  26. Appleyard MV, Murray KE, Coates PJ, Wullschleger S, Bray SE, Kernohan NM et al. Phenformin as prophylaxis and therapy in breast cancer xenografts. Br J Cancer 2012; 106: 1117–1122.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Pollak M . Potential applications for biguanides in oncology. J Clin Invest 2013; 123: 3693–3700.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Corton JM, Gillespie JG, Hawley SA, Hardie DG . 5-aminoimidazole-4-carboxamide ribonucleoside A specific method for activating AMP-activated protein kinase in intact cells? Eur J Biochem 1995; 229: 558–565.

    Article  CAS  PubMed  Google Scholar 

  29. El-Masry OS, Brown BL, Dobson PR . Effects of activation of AMPK on human breast cancer cell lines with different genetic backgrounds. Oncol Lett 2012; 3: 224–228.

    Article  CAS  PubMed  Google Scholar 

  30. Rosilio C, Lounnas N, Nebout M, Imbert V, Hagenbeek T, Spits H et al. The metabolic perturbators metformin, phenformin and AICAR interfere with the growth and survival of murine PTEN-deficient T cell lymphomas and human T-ALL/T-LL cancer cells. Cancer Lett 2013; 336: 114–126.

    Article  CAS  PubMed  Google Scholar 

  31. Cool B, Zinker B, Chiou W, Kifle L, Cao N, Perham M et al. Identification and characterization of a small molecule AMPK activator that treats key components of type 2 diabetes and the metabolic syndrome. Cell Metab 2006; 3: 403–416.

    Article  CAS  PubMed  Google Scholar 

  32. Hawley SA, Fullerton MD, Ross FA, Schertzer JD, Chevtzoff C, Walker KJ et al. The ancient drug salicylate directly activates AMP-activated protein kinase. Science 2013; 336: 918–922.

    Article  Google Scholar 

  33. Elder DJ, Hague A, Hicks DJ, Paraskeva C . Differential growth inhibition by the aspirin metabolite salicylate in human colorectal tumor cell lines: enhanced apoptosis in carcinoma and in vitro-transformed adenoma relative to adenoma relative to adenoma cell lines. Cancer Res 1996; 56: 2273–2276.

    CAS  PubMed  Google Scholar 

  34. Hardie DG . AMPK: a target for drugs and natural products with effects on both diabetes and cancer. Diabetes 2013; 62: 2164–2172.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Liang J, Shao SH, Xu ZX, Hennessy B, Ding Z, Larrea M et al. The energy sensing LKB1-AMPK pathway regulates p27(kip1) phosphorylation mediating the decision to enter autophagy or apoptosis. Nat Cell Biol 2007; 9: 218–224.

    Article  CAS  PubMed  Google Scholar 

  36. Bungard D, Fuerth BJ, Zeng PY, Faubert B, Maas NL, Viollet B et al. Signaling kinase AMPK activates stress-promoted transcription via histone H2B phosphorylation. Science 2010; 329: 1201–1205.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Zannella VE, Cojocari D, Hilgendorf S, Vellanki RN, Chung S, Wouters BG et al. AMPK regulates metabolism and survival in response to ionizing radiation. Radiother Oncol 2011; 99: 293–299.

    Article  CAS  PubMed  Google Scholar 

  38. Vander Heiden MG, Cantley LC, Thompson CB . Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 2009; 324: 1029–1033.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Liang J, Mills G . AMPK: a contextual oncogene or tumor suppressor? Cancer Res 2013; 73: 2929–2935.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Pachman L, Esterly N, Peterson R . The effect of salicylate on the metabolism of normal and stimulated human lymphocytes in vitro. J Clin Invest 1971; 50: 226–230.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Brody T . Action of sodium salicylate and related compounds on tissue metabolism in vitro. J Pharmacol Exp Ther 1956; 117: 39–51.

    CAS  PubMed  Google Scholar 

  42. Nicholls DG . Spare respiratory capacity, oxidative stress and excitotoxicity. Biochem Soc Trans 2009; 37: 1385–1388.

    Article  CAS  PubMed  Google Scholar 

  43. Hawley SA, Pan DA, Mustard KJ, Ross L, Bain J, Edelman AM et al. Calmodulin-dependent protein kinase kinase-beta is an alternative upstream kinase for AMP-activated protein kinase. Cell Metab 2005; 2: 9–19.

    Article  CAS  PubMed  Google Scholar 

  44. Goransson O, McBride A, Hawley SA, Ross FA, Shpiro N, Foretz M et al. Mechanism of action of A-769662, a valuable tool for activation of AMP-activated protein kinase. J Biol Chem 2007; 282: 32549–32560.

    Article  PubMed  Google Scholar 

  45. Kalender A, Selvaraj A, Kim SY, Gulati P, Brule S, Viollet B et al. Metformin, independent of AMPK, inhibits mTORC1 in a rag GTPase-dependent manner. Cell Metab 2010; 11: 390–401.

    CAS  PubMed  PubMed Central  Google Scholar 

  46. Zakikhani M, Dowling R, Fantus IG, Sonenberg N, Pollak M . Metformin is an AMP kinase-dependent growth inhibitor for breast cancer cells. Cancer Res 2006; 66: 10269–10273.

    Article  CAS  PubMed  Google Scholar 

  47. Dykens JA, Jamieson J, Marroquin L, Nadanaciva S, Billis PA, Will Y . Biguanide-induced mitochondrial dysfunction yields increased lactate production and cytotoxicity of aerobically-poised HepG2 cells and human hepatocytes in vitro. Toxicol Appl Pharmacol 2008; 233: 203–210.

    Article  CAS  PubMed  Google Scholar 

  48. Shu Y, Sheardown SA, Brown C, Owen RP, Zhang S, Castro RA et al. Effect of genetic variation in the organic cation transporter 1 (OCT1) on metformin action. J Clin Invest 2007; 117: 1422–1431.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Zadra G, Photopoulos C, Tyekucheva S, Heidari P, Weng QP, Fedele G et al. A novel direct activator of AMPK inhibits prostate cancer growth by blocking lipogenesis. EMBO Mol Med 2014; 6: 519–538.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Huang X, Wullschleger S, Shpiro N, McGuire VA, Sakamoto K, Woods YL et al. Important role of the LKB1-AMPK pathway in suppressing tumorigenesis in PTEN-deficient mice. Biochem J 2008; 412: 211–221.

    Article  CAS  PubMed  Google Scholar 

  51. Shackelford DB, Abt E, Gerken L, Vasquez DS, Seki A, Leblanc M et al. LKB1 inactivation dictates therapeutic response of non-small cell lung cancer to the metabolism drug phenformin. Cancer Cell 2013; 23: 143–158.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Faubert B, Vincent EE, Poffenberger MC, Jones RG . The AMP-activated protein kinase (AMPK) and cancer: many faces of a metabolic regulator. Cancer letters 2014.

  53. Garber K . Energy deregulation: licensing tumors to grow. Science 2006; 312: 1158–1159.

    Article  CAS  PubMed  Google Scholar 

  54. Guigas B, Bertrand L, Taleux N, Foretz M, Wiernsperger N, Vertommen D et al. 5-Aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside and metformin inhibit hepatic glucose phosphorylation by an AMP-activated protein kinase-independent effect on glucokinase translocation. Diabetes 2006; 55: 865–874.

    Article  CAS  PubMed  Google Scholar 

  55. Vincent MF, Bontemps F, Van den Berghe G . Inhibition of glycolysis by 5-amino-4-imidazolecarboxamide riboside in isolated rat hepatocytes. Biochem J 1992; 281 (Pt 1): 267–272.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Morita M, Gravel SP, Chenard V, Sikstrom K, Zheng L, Alain T et al. mTORC1 controls mitochondrial activity and biogenesis through 4E-BP-dependent translational regulation. Cell Metab 2013; 18: 698–711.

    Article  CAS  PubMed  Google Scholar 

  57. Brand MD, Nicholls DG . Assessing mitochondrial dysfunction in cells. Biochem J 2011; 435: 297–312.

    Article  CAS  PubMed  Google Scholar 

  58. Jeon SM, Chandel NS, Hay N . AMPK regulates NADPH homeostasis to promote tumour cell survival during energy stress. Nature 2012; 485: 661–665.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Godlewski J, Nowicki MO, Bronisz A, Nuovo G, Palatini J, De Lay M et al. MicroRNA-451 regulates LKB1/AMPK signaling and allows adaptation to metabolic stress in glioma cells. Mol Cell 2010; 37: 620–632.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Chhipa RR, Wu Y, Mohler JL, Ip C . Survival advantage of AMPK activation to androgen-independent prostate cancer cells during energy stress. Cell Signal 2010; 22: 1554–1561.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  61. Laderoute KR, Amin K, Calaoagan JM, Knapp M, Le T, Orduna J et al. 5′-AMP-activated protein kinase (AMPK) is induced by low-oxygen and glucose deprivation conditions found in solid-tumor microenvironments. Mol Cell Biol 2006; 26: 5336–5347.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  62. Crissman HA, Steinkamp JA . simultaneous measurement of DNA, protein, and cell volume in single cells from large mammalian cell populations. J Cell Biol 1973; 59: 766–771.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  63. Vincent EE, Elder DJ, Curwen J, Kilgour E, Hers I, Tavare JM . Targeting non-small cell lung cancer cells by dual inhibition of the insulin receptor and the insulin-like growth factor-1 receptor. PLoS ONE 2013; 8: e66963.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  64. Faubert B, Vincent EE, Griss T, Samborska B, Izreig S, Svensson RU et al. Loss of the tumor suppressor LKB1 promotes metabolic reprogramming of cancer cells via HIF-1alpha. Proc Natl Acad Sci USA 2014; 111: 2554–2559.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We acknowledge Douglas Elder and Nicole Beauchemin, and members of the Jones Laboratory for critical reading of this manuscript. EEV, PPC and TG were funded by the McGill Integrated Cancer Research Training Program (MICRTP) and JB was funded by Fonds de researche Santé Québec. This work was supported by grants to RGJ from the Canadian Institute Health Research (CIHR) (MOP-93799), Canadian Cancer Society (2010-700586) and Terry Fox Research Foundation (TEF-116128). This work is dedicated to the memory of Rosalind Goodman.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to R G Jones.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Vincent, E., Coelho, P., Blagih, J. et al. Differential effects of AMPK agonists on cell growth and metabolism. Oncogene 34, 3627–3639 (2015). https://doi.org/10.1038/onc.2014.301

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2014.301

This article is cited by

Search

Quick links