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

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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.

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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

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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.

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Correspondence to Manju Ray.

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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.

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Handling Editor: T. Wallimann and R. Harris.

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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

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