Skip to main content

Advertisement

SpringerLink
Log in

Creatine supplementation with methylglyoxal: a potent therapy for cancer in experimental models

  • Original Article
  • Published:
Amino Acids Aims and scope Submit manuscript

Abstract

The anti-cancer effect of methylglyoxal (MG) is now well established in the literature. The main aim of this study was to investigate the effect of creatine as a supplement in combination with MG both in vitro and in vivo. In case of the in vitro studies, two different cell lines, namely MCF-7 (human breast cancer cell line) and C2C12 (mouse myoblast cell line) were chosen. MG in combination with creatine showed enhanced apoptosis as well as higher cytotoxicity in the breast cancer MCF-7 cell line, compared to MG alone. Pre-treatment of well-differentiated C2C12 myotubes with cancerogenic 3-methylcholanthrene (3MC) induced a dedifferentiation of these myotubes towards cancerous cells (that mimic the effect of 3MC observed in solid fibro-sarcoma animal models) and subsequent exposure of these induced cancer cells with MG proved to be cytotoxic. Thus, creatine plus ascorbic acid enhanced the anti-cancer effects of MG. In contrast, when normal C2C12 muscle cells or myotubes (mouse normal myoblast cell line) were treated with MG or MG plus creatine and ascorbic acid, no detrimental effects were seen. This indicated that cytotoxic effects of MG are specifically limited towards cancer cells and are further enhanced when MG is used in combination with creatine and ascorbic acid. For the in vivo studies, tumors were induced by injecting Sarcoma-180 cells (2 × 106 cells/mouse) in the left hind leg. After 7 days of tumor inoculation, treatments were started with MG (20 mg/kg body wt/day, via the intravenous route), with or without creatine (150 mg/kg body wt/day, fed orally) and ascorbic acid (50 mg/kg body wt/day, fed orally) and continued for 10 consecutive days. Significant regression of tumor size was observed when Sarcoma-180 tumor-bearing mice were treated with MG and even more so with the aforesaid combination. The creatine-supplemented group demonstrated better overall survival in comparison with tumor-bearing mice without creatine. In conclusion, it may be stated that the anti-cancer effect of MG is enhanced by concomitant creatine supplementation, both in chemically transformed (by 3MC) muscle cells in vitro as well as in sarcoma animal model in vivo. These data strongly suggest that creatine supplementation may gain importance as a safe and effective supplement in therapeutic intervention with the anti-cancer agent MG.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

CK:

Creatine kinase

MCK:

Muscle-specific cytosolic CK

BCK:

Brain-specific cytosolic CK

MtCK:

Mitochondrial CK

3MC:

3-Methylcholanthrene

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

EAC:

Ehrlich ascites carcinoma

MTT:

(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)

DMSO:

Dimethylsulfoxide

References

  • Albini A, Pennesi G, Donatelli F, Cammarota R, De Flora S, Noonan DM (2010) Cardiotoxicity of anticancer drugs: the need for cardio-oncology and cardio-oncological prevention. J Natl Cancer Inst 102:14–25

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bagui S, Ray M, Ray S (1999) Glyceraldehyde-3-phosphate dehydrogenase from Ehrlich ascites carcinoma cells. Its possible role in the high glycolysis of malignant cells. Eur J Biochem 262:386–395

    Article  CAS  PubMed  Google Scholar 

  • Balasubramani M, Day BW, Schoen RE, Getzenberg RH (2006) Altered expression and localization of creatine kinase B, heterogeneous nuclear ribonucleoprotein F, and high mobility group box 1 protein in the nuclear matrix associated with colon cancer. Cancer Res 66:763–769

    Article  CAS  PubMed  Google Scholar 

  • Biswas S, Ray M, Misra S, Dutta DP, Ray S (1997) Selective inhibition of mitochondrial respiration and glycolysis in human leukaemic leukocytes by methylglyoxal. Biochem J 323:343–348

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Carney DN, Zweig MH, Ihde DC, Cohen MH, Makuch RW, Gazdar AF (1984) Elevated serum creatine kinase BB levels in patients with small cell lung cancer. Cancer Res 44:5399–5403

    CAS  PubMed  Google Scholar 

  • Chen Qi, Espey MG, Krishna MC, Mitchell JB, Corpe CP, Buettner GR, Shacter E, Levine M (2005) Pharmacologic ascorbic acid concentrations selectively kill cancer cells: action as a pro-drug to deliver hydrogen peroxide to tissues. Proc Natl Acad Sci 102(38):13604–13609

    Article  Google Scholar 

  • Das MR, Bag AK, Saha S, Ghosh A, Dey SK, Das P, Mandal C, Ray S, Chakrabarti S, Ray M, Jana SS (2016) Molecular association of Glucose-6-phosphate isomerase and Pyruvate kinase M2 with Glyceraldehyde-3-phosphate dehydrogenase in cancer cells. BMC Cancer 16(1):152

    Article  PubMed  PubMed Central  Google Scholar 

  • Du J, Cullen JJ, Buettner GR (2012) Ascorbic acid: chemistry, biology and the treatment of cancer. Biochem Biophys Acta 1826(2):443–457

    CAS  PubMed  PubMed Central  Google Scholar 

  • Együd LG, Szent-Györgyi A (1968) Cancerostatic action of methylglyoxal. Science 160:1140

    Article  PubMed  Google Scholar 

  • Ferreira-da-Silva F, Pereira PJ, Gales L, Roessle M, Svergun DI, Moradas-Ferreira P, Damas AM (2006) The crystal and solution structures of glyceraldehyde-3-phosphate dehydrogenase reveal different quaternary structures. J Biol Chem 281:33433–33440

    Article  CAS  PubMed  Google Scholar 

  • Gazdar AF, Zweig MH, Carney DN, Van Steirteghen AC, Baylin SB, Minna JD (1981) Levels of creatine kinase and its BB isoenzyme in lung cancer specimens and cultures. Cancer Res 41:2773–2777

    CAS  PubMed  Google Scholar 

  • Ghosh M, Talukdar D, Ghosh S, Bhattacharyya N, Ray M, Ray S (2006) In vivo assessment of toxicity and pharmacokinetics of methylglyoxal. Augmentation of the curative effect of methylglyoxal on cancer-bearing mice by ascorbic acid and creatine. Toxicol Appl Pharmacol 212:45–58

    Article  CAS  PubMed  Google Scholar 

  • Ghosh A, Bera S, Ghosal S, Ray S, Basu A, Ray M (2011) Differential inhibition/inactivation of mitochondrial complex I implicates its alteration in malignant cells. Biochem Moscow 76:1051–1060

    Article  CAS  Google Scholar 

  • Halder J, Ray M, Ray S (1993) Inhibition of glycolysis and mitochondrial respiration of Ehrlich ascites carcinoma cells by methylglyoxal. Int J Cancer 54:443–449

    Article  CAS  PubMed  Google Scholar 

  • Horska A, Ulug AM, Melhem ER, Filippi CG, Burger PC, Edgar MA, Souweidane MM, Carson BS, Barker PB (2001) Proton magnetic resonance spectroscopy of choroid plexus tumors in children. J Magn Reson Imaging 14:78–82

    Article  CAS  PubMed  Google Scholar 

  • Ishiguro Y, Kato K, Akatsuska H, Ito T (1990) The diagnostic and prognostic value of pretreatment serum creatine kinase BB levels in patients with neuroblastoma. Cancer 65:2014–2019

    Article  CAS  PubMed  Google Scholar 

  • Kinoshita Y, Yokota A (1997) Absolute concentrations of metabolites in human brain tumors using in vitro proton magnetic resonance spectroscopy. NMR Biomed 10:2–12

    Article  CAS  PubMed  Google Scholar 

  • Kornacker M, Schlattner U, Wallimann T, Verneris MR, Negrin RS, Kornacker B, Staratschek-Jox A, Diehl V, Wolf J (2001) Hodgkin disease-derived cell lines expressing ubiquitous mitochondrial creatine kinase show growth inhibition by cyclocreatine treatment independent of apoptosis. Int J Cancer 94:513–519

    Article  CAS  PubMed  Google Scholar 

  • Kristensen CA, Askenasy N, Jain RK, Koretsky AP (1999) Creatine and cyclocreatine treatment of human colon adenocarcinoma xenografts: 31P and 1H magnetic resonance spectroscopic studies. Br J Cancer 79:278–285

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lawler JM, Barnes WS, Wu G, Song W, Demaree S (2002) Direct antioxidant properties of creatine. Biochem Biophys Res Commun 290:47–52

    Article  CAS  PubMed  Google Scholar 

  • Lehnhardt FG, Bock C, Rohn G, Ernestus RI, Hoehn M (2005) Metabolic differences between primary and recurrent human brain tumors: a 1H NMR spectroscopic investigation. NMR Biomed 18:371–382

    Article  CAS  PubMed  Google Scholar 

  • Loo JM, Scherl A, Nguyen A, Man FY, Weinberg E, Zeng Z, Saltz L, Paty PB, Tavazoie SF (2015) Extracellular metabolic energetics can promote cancer progression. Cell 160:393–406

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Meffert G, Gellerich FN, Margreiter R, Wyss M (2005) Elevated creatine kinase activity in primary hepatocellular carcinoma. BMC Gastroenterol 5:9

    Article  PubMed  PubMed Central  Google Scholar 

  • Miller EE, Evans AE, Cohn M (1993) Inhibition of rate of tumor growth by creatine and cyclocreatine. Proc Natl Acad Sci USA 90:3304–3308

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63

    Article  CAS  PubMed  Google Scholar 

  • Mostafa G, Lamont C, Greene FL (2006) Review of Surgery: Basic Science and Clinical Topics for ABSITE. New York, Springer Science and Business Media. 37–38. ISBN 978-0-387-44952-4

  • Patra S, Bera S, Sinha Roy S, Ghoshal S, Ray S, Basu A, Schlattner U, Wallimann T, Ray M (2008) Progressive decrease of phosphor creatine, creatine and creatine kinase in skeletal muscle upon transformation to sarcoma. FEBS J 275:3236–3247

    Article  CAS  PubMed  Google Scholar 

  • Patra S, Ghosh S, Bera S, Roy A, Ray S, Ray M (2009) Molecular characterization of tumor associated glyceraldehyde-3-phosphate dehydrogenase. Biochem Moscow 74:717–727

    Article  CAS  Google Scholar 

  • Patra S, Ghosh A, Roy SS, Bera S, Das M, Talukdar D, Ray S, Wallimann T, Ray M (2012) A short review on creatine–creatine kinase system in relation to cancer and some experimental results on creatine as adjuvant in cancer therapy. Amino Acids 42:2319–2330

    Article  CAS  PubMed  Google Scholar 

  • Payne RM, Haas RC, Strauss AW (1991) Structural characterization and tissue specific expression of them RNAs encoding isoenzymes from two rat mitochondrial creatine kinase genes. Biochem Biophys Acta 1089:352–361

    CAS  PubMed  Google Scholar 

  • Ray M, Ray S (1998) Methylglyoxal: from putative intermediate of glucose breakdown to its role in understanding that excessive ATP formation in cells may lead to malignancy. Curr Sci 75:103–113

    CAS  Google Scholar 

  • Ray M, Halder J, Dutta SK, Ray S (1991) Inhibition of respiration of tumor cells by methylglyoxal and protection of inhibition by lactaldehyde. Int J Cancer 47:603–609

    Article  CAS  PubMed  Google Scholar 

  • Ray S, Biswas S, Ray M (1997a) Similar nature of inhibition of mitochondrial respiration of heart tissue and malignant cells by methylglyoxal. A vital clue to understand the biochemical basis of malignancy. Mol Cell Biochem 171:95–103

    Article  CAS  PubMed  Google Scholar 

  • Ray M, Basu N, Ray S (1997b) Inactivation of glyceraldehyde-3-phosphate dehydrogenase of human malignant cells by methylglyoxal. Mol Cell Biochem 177:21–26

    Article  CAS  PubMed  Google Scholar 

  • Roy SS, Biswas S, Ray M, Ray S (2003) Protective effect of creatine against inhibition by methylglyoxal of mitochondrial respiration of cardiac cells. Biochem J 372:661–669

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Saha S, Dey SK, Das P, Jana SS (2011) Increased expression of nonmuscle myosin IIs is associated with 3MC-induced mouse tumor. FEBS J 278(21):4025–4034

    Article  CAS  PubMed  Google Scholar 

  • Santacruz L, Darrabie MD, Mantilla JG, Mishra R, Feger BJ, Jacobs DO (2015) Creatine supplementation reduces doxorubicin-induced cardiomyocellular injury. Cardiovasc Toxicol 15:180–188

    Article  CAS  PubMed  Google Scholar 

  • Sestili P, Martinelli C, Colombo E, Barbieri E, Potenza L, Sartini S, Fimognari C (2011) Creatine as an antioxidant. Amino Acids 40:1385–1396

    Article  CAS  PubMed  Google Scholar 

  • Sullivan WJ, Christofk HR (2015) The metabolic milieu of metastases. Cell 160:363–364

    Article  CAS  PubMed  Google Scholar 

  • Szent-Györgi A (1978) The living state and cancer. Ciba Found Symp 67:3–18

    Google Scholar 

  • Tokarska-Schlattner M, Zaugg M, Zuppinger C, Wallimann T, Schlattner U (2006) New insights into doxorubicin-induced cardiotoxicity: the critical role of cellular energetics. J Mol Cell Cardiol 41:389–405

    Article  CAS  PubMed  Google Scholar 

  • Tuma DJ, Donohue TM Jr, Medina VA, Sorrell MF (1984) Enhancement of acetaldehyde-protein adduct formation by L-ascorbate. Arch Biochem Biophy 234:377–381

    Article  CAS  Google Scholar 

  • Van Pilsum JF, Stephens GC, Taylor D (1972) Distribution of creatine, guanidinoacetate and the enzymes for their biosynthesis in the animal kingdom. Biochem J 126:325–345

    Article  PubMed Central  Google Scholar 

  • Wallimann T, Wyss M, Brdiczka D, Nicolay K, Eppenberger HM (1992) Intracellular compartmentation, structure and function of creatine kinase isozymes in tissues with high and fluctuating energy demands: the ‘phosphocreatine’ circuit for cellular energy homeostasis. Biochem J 281:21–40

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wallimann T, Tokarska-Schlattner M, Neumann D, Epand RM, Epand RF, Andres RH, Widmer HR, Saks VA, Agarkova I, Schlattner U (2007) The phosphocreatine circuit: molecular and cellular physiology of creatine kinases, sensitivity to free radicals, and enhancement by creatine supplementation. In: Saks VA (ed) Molecular Systems Bioenergetics, chap 7. Wiley, pp 195–265. doi:10.1002/9783527621095

  • Wallimann T, Tokarska-Schlattner M, Schlattner U (2011) The creatine kinase system and pleiotropic effects of creatine. Amino Acids 40:1271–1296

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wyss M, Kaddurah-Daouk R (2000) Creatine and creatinine metabolism. Physiol Rev 80:1107–1213

    CAS  PubMed  Google Scholar 

  • Zarghami N, Giai M, Yu H, Roagna R, Ponzone R, Katsaros D, Sismondi P, Diamandis EP (1996) Creatine kinase BB isoenzyme levels in tumour cytosols and survival of breast cancer patients. Br J Cancer 73:386–390

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This research is funded by the Council of Scientific and Industrial Research (CSIR), Government of India; University Grants Commission (UGC), Government of India, and Department of Science and Technology (DST) Nanomission, Government of India. We thank Dr. Alok Ghosh for his help in some experiments. The authors are indebted to late Dr. Subhankar Ray who was intimately associated to the work and left us in June 2015. We dedicate this manuscript to him.

Author information

Authors and Affiliations

  1. Department of Biophysics, Bose Institute, P 1/12, CIT Road, Scheme VIIM, Kolkata, 700 054, West Bengal, India

    Aparajita Pal, Anirban Roy & Manju Ray

Authors
  1. Aparajita Pal
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anirban Roy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Manju Ray
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Manju Ray.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest in this work.

Ethical approval

All procedures performed in studies involving animals were in accordance with the ethical standards of the Institutional Animal Ethics Committee of Bose Institute, Kolkata, India.

Additional information

Handling Editor: T. Wallimann and R. Harris.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Pal, A., Roy, A. & Ray, M. Creatine supplementation with methylglyoxal: a potent therapy for cancer in experimental models. Amino Acids 48, 2003–2013 (2016). https://doi.org/10.1007/s00726-016-2224-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00726-016-2224-1

Keywords

Search

Navigation