Reversal of the glycolytic phenotype by dichloroacetate inhibits metastatic breast cancer cell growth in vitro and in vivo

Abstract

The glycolytic phenotype is a widespread phenomenon in solid cancer forms, including breast cancer. Dichloroacetate (DCA) has recently been proposed as a novel and relatively non-toxic anti-cancer agent that can reverse the glycolytic phenotype in cancer cells through the inhibition of pyruvate dehydrogenase kinase. We have examined the effect of DCA against breast cancer cells, including in a highly metastatic in vivo model. The growth of several breast cancer cell lines was found to be inhibited by DCA in vitro. Further examination of 13762 MAT rat mammary adenocarcinoma cells found that reversal of the glycolytic phenotype by DCA correlated with the inhibition of proliferation without any increase in cell death. This was despite a small but significant increase in caspase 3/7 activity, which may sensitize cancer cells to other apoptotic triggers. In vivo, DCA caused a 58% reduction in the number of lung metastases observed macroscopically after injection of 13762 MAT cells into the tail vein of rats (P = 0.0001, n ≥ 9 per group). These results demonstrate that DCA has anti-proliferative properties in addition to pro-apoptotic properties, and can be effective against highly metastatic disease in vivo, highlighting its potential for clinical use.

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References

  1. 1.

    Fantin VR, St-Pierre J, Leder P (2006) Attenuation of LDH—a expression uncovers a link between glycolysis, mitochondrial physiology, and tumor maintenance. Cancer Cell 9:425–434

    Article  CAS  PubMed  Google Scholar 

  2. 2.

    Gatenby R, Gillies RJ (2007) Glycolysis in cancer: a potential target for therapy. Int J Biochem Cell Biol 39:1358–1366

    Article  CAS  PubMed  Google Scholar 

  3. 3.

    Isidoro A, Martnez M, Fernndez PL, Ortega AD, Santamara G, Chamorro M, Reed JC, Cuezva JM (2004) Alteration of the bioenergetic phenotype of mitochondria is a hallmark of breast, gastric, lung and oesophageal cancer. Biochem J 378:17–20

    Article  CAS  PubMed  Google Scholar 

  4. 4.

    Isidoro A, Casado E, Redondo A, Acebo P, Espinosa E, Alonso AM, Cejas P, Hardisson D, Fresno Vara JA, Belda-Iniesta C, Gonzlez-Barn M, Cuezva JM (2005) Breast carcinomas fulfill the Warburg hypothesis and provide metabolic markers of cancer prognosis. Carcinogenesis 26:2095–2104

    Article  CAS  PubMed  Google Scholar 

  5. 5.

    Robey IF, Lien AD, Welsh SJ, Baggett BK, Gillies RJ (2005) Hypoxia-inducible factor-1α and the glycolytic phenotype in tumors. Neoplasia 7:324–330

    Article  CAS  PubMed  Google Scholar 

  6. 6.

    Gatenby RA, Smallbone K, Maini PK, Rose F, Averill J, Nagle RB, Worrall L, Gillies RJ (2007) Cellular adaptations to hypoxia and acidosis during somatic evolution of breast cancer. Br J Cancer 97:646–653

    Article  CAS  PubMed  Google Scholar 

  7. 7.

    Gudi R, Bowker-Kinley MM, Kedishvili NY, Zhao Y, Popov KM (1995) Diversity of the pyruvate dehydrogenase kinase gene family in humans. J Biol Chem 270:28989–28994

    Article  CAS  PubMed  Google Scholar 

  8. 8.

    Bonnet S, Archer SL, Allalunis-Turner J, Haromy A, Beaulieu C, Thompson R, Lee CT, Lopaschuk GD, Puttagunta L, Bonnet S, Harry G, Hashimoto K, Porter CJ, Andrade MA, Thebaud B, Michelakis ED (2007) A mitochondria-K+ channel axis is suppressed in cancer and its normalization promotes apoptosis and inhibits cancer growth. Cancer Cell 11:37–51

    Article  CAS  PubMed  Google Scholar 

  9. 9.

    Wong JY, Huggins GS, Debidda M, Munshi NC, Vivo ID (2008) Dichloroacetate induces apoptosis in endometrial cancer cells. Gynecol Oncol 109:394–402

    Article  CAS  PubMed  Google Scholar 

  10. 10.

    Kaufmann P, Engelstad K, Wei Y, Jhung S, Sano MC, Shungu DC, Millar WS, Hong X, Gooch CL, Mao X, Pascual JM, Hirano M, Stacpoole PW, DiMauro S, Vivo DCD (2006) Dichloroacetate causes toxic neuropathy in MELAS: a randomized, controlled clinical trial. Neurology 66:324–330

    Article  CAS  PubMed  Google Scholar 

  11. 11.

    Stacpoole PW, Kerr DS, Barnes C, Bunch ST, Carney PR, Fennell EM, Felitsyn NM, Gilmore RL, Greer M, Henderson GN, Hutson AD, Neiberger RE, O’Brien RG, Perkins LA, Quisling RG, Shroads AL, Shuster JJ, Silverstein JH, Theriaque DW, Valenstein E (2006) Controlled clinical trial of dichloroacetate for treatment of congenital lactic acidosis in children. Pediatrics 117:1519–1531

    Article  PubMed  Google Scholar 

  12. 12.

    Pearson H (2007) Cancer patients opt for unapproved drug. Nature 446:474–475

    Article  CAS  PubMed  Google Scholar 

  13. 13.

    Parish CR, Freeman C, Brown KJ, Francis DJ, Cowden WB (1999) Identification of sulfated oligosaccharide-based inhibitors of tumor growth and metastasis using novel in vitro assays for angiogenesis and heparanase activity. Cancer Res 59:3433–3441

    CAS  PubMed  Google Scholar 

  14. 14.

    Blackburn AC, McLary SC, Naeem R, Luszcz J, Stockton DW, Donehower LA, Mohammed M, Mailhes JB, Soferr T, Naber SP, Otis CN, Jerry DJ (2004) Loss of heterozygosity occurs via mitotic recombination in Trp53+/− mice and associates with mammary tumor susceptibility of the BALB/c strain. Cancer Res 64:5140–5147

    Article  CAS  PubMed  Google Scholar 

  15. 15.

    Schmuck E, Cappello J, Coggan M, Brew J, Cavanaugh JA, Blackburn AC, Baker RT, Eyre HJ, Sutherland GR, Board PG (2008) Deletion of Glu155 causes a deficiency of glutathione transferase Omega 1-1 but does not alter sensitivity to arsenic trioxide and other cytotoxic drugs. Int J Biochem Cell Biol 40:2553–2559

    Article  CAS  PubMed  Google Scholar 

  16. 16.

    Quah BJ, Warren HS, Parish CR (2007) Monitoring lymphocyte proliferation in vitro and in vivo with the intracellular fluorescent dye carboxyfluorescein diacetate succinimidyl ester. Nat Protoc 2:2049–2056

    Article  CAS  PubMed  Google Scholar 

  17. 17.

    Beutler E (1975) Red cell metabolism: a manual of biochemical methods, 2nd edn. Grune & Stratton Inc., New York

  18. 18.

    Saghir SA, Schultz IR (2002) Low-dose pharmacokinetics and oral bioavailability of dichloroacetate in naive and GST-zeta-depleted rats. Environ Health Perspect 110:757–763

    CAS  PubMed  Google Scholar 

  19. 19.

    Gonzalez-Leon A, Schultz IR, Xu G, Bull RJ (1997) Pharmacokinetics and metabolism of dichloroacetate in the F344 rat after prior administration in drinking water. Toxicol Appl Pharmacol 146:189–195

    Article  CAS  PubMed  Google Scholar 

  20. 20.

    Board PG, Anders MW (2005) Human glutathione transferase zeta. Methods Enzymol 401:61–77

    Article  CAS  PubMed  Google Scholar 

  21. 21.

    Lim CEL, Matthaei KI, Blackburn AC, Davis RP, Dahlstrom JE, Koina ME, Anders MW, Board PG (2004) Mice deficient in glutathione transferase zeta/maleylacetoacetate isomerase exhibit a range of pathological changes and elevated expression of alpha, mu and pi class glutathione transferases. Am J Pathol 165:679–693

    CAS  PubMed  Google Scholar 

  22. 22.

    Cao W, Yacoub S, Shiverick KT, Namiki K, Sakai Y, Porvasnik S, Urbanek C, Rosser CJ (2008) Dichloroacetate (DCA) sensitizes both wild-type and over expressing Bcl-2 prostate cancer cells in vitro to radiation. Prostate 68:1223–1231

    Article  CAS  PubMed  Google Scholar 

  23. 23.

    Bowker-Kinley MM, Davis WI, Wu P, Harris RA, Popov KM (1998) Evidence for existence of tissue-specific regulation of the mammalian pyruvate dehydrogenase complex. Biochem J 329:191–196

    CAS  PubMed  Google Scholar 

  24. 24.

    Lu CW, Lin SC, Chen KF, Lai YY, Tsai SJ (2008) Induction of pyruvate dehydrogenase kinase-3 by hypoxia-inducible factor-1 promotes metabolic switch and drug resistance. J Biol Chem 283:28106–28114

    Article  CAS  PubMed  Google Scholar 

  25. 25.

    DiPietrantonio AM, Hsieh T, Wu JM (1999) Activation of caspase 3 in HL-60 cells exposed to hydrogen peroxide. Biochem Biophys Res Commun 255:477–482

    Article  CAS  PubMed  Google Scholar 

  26. 26.

    Fischer K, Hoffmann P, Voelkl S, Meidenbauer N, Ammer J, Edinger M, Gottfried E, Schwarz S, Rothe G, Hoves S, Renner K, Timischl B, Mackensen A, Kunz-Schughart L, Andreesen R, Krause SW, Kreutz M (2007) Inhibitory effect of tumor cell-derived lactic acid on human T cells. Blood 109:3812–3819

    Article  CAS  PubMed  Google Scholar 

  27. 27.

    Mori M, Yamagata T, Goto T, Saito S, Momoi MY (2004) Dichloroacetate treatment for mitochondrial cytopathy: long-term effects in MELAS. Brain Dev 26:453–458

    Article  PubMed  Google Scholar 

  28. 28.

    Shangraw RE, Lohan-Mannion D, Hayes A, Moriarty RM, Fu R, Robinson ST (2008) Dichloroacetate stabilizes the intraoperative acid-base balance during liver transplantation. Liver Transpl 14:989–998

    Article  PubMed  Google Scholar 

  29. 29.

    Stacpoole PW, Gilbert LR, Neiberger RE, Carney PR, Valenstein E, Theriaque DW, Shuster JJ (2008) Evaluation of long-term treatment of children with congenital lactic acidosis with dichloroacetate. Pediatrics 121:e1223–e1228

    Article  PubMed  Google Scholar 

  30. 30.

    Theodoratos A, Tu WJ, Cappello J, Blackburn AC, Matthaei K, Board PG (2009) Phenylalanine-induced leucopenia in genetic and dichloroacetic acid generated deficiency of glutathione transferase zeta. Biochem Pharmacol 77:1358–1363

    Article  CAS  PubMed  Google Scholar 

  31. 31.

    Mayers RM, Leighton B, Kilgour E (2005) PDH kinase inhibitors: a novel therapy for Type II diabetes? Biochem Soc Trans 33:367–370

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This research was supported by a grant from the National Breast Cancer Foundation Australia, and by NHMRC 366787 R.D. Wright Career Development Award.

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Correspondence to Anneke C. Blackburn.

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Sun, R.C., Fadia, M., Dahlstrom, J.E. et al. Reversal of the glycolytic phenotype by dichloroacetate inhibits metastatic breast cancer cell growth in vitro and in vivo. Breast Cancer Res Treat 120, 253–260 (2010). https://doi.org/10.1007/s10549-009-0435-9

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Keywords

  • Dichloroacetate
  • Breast cancer
  • Glycolysis
  • Metastasis
  • Animal model