Journal of Cancer Research and Treatment
ISSN (Print): 2374-1996 ISSN (Online): 2374-2003 Website: https://www.sciepub.com/journal/jcrt Editor-in-chief: Jean Rommelaere
Open Access
Journal Browser
Go
Journal of Cancer Research and Treatment. 2018, 6(1), 18-24
DOI: 10.12691/jcrt-6-1-4
Open AccessArticle

The Antioxidant Glycolysis Inhibitor (Citric Acid) Induces a Dose-dependent Caspase-mediated Apoptosis and Necrosis in Glioma Cells

Salah Mohamed El Sayed1, 2, , Hussam H. Baghdadi1, Nassar Ayoub Abdellatif Omar3, 4, Amal Nor Edeen Ahmad Allithy5, 6, Nahed Mohammed Hablas7, Ahmed Ragab Fakhreldin8, Reham Abelsalam Mariah1, 9, Momen El-Shazley10, 11, Mongi Ayat1, Sayed Mostafa12, Mostafa Abu-el Naga13, 14 and Mohamed Abdel-Halim15

1Department of Biochemistry and Molecular Medicine, Taibah Faculty of Medicine, Taibah University, Al-Madinah Al-Munawwarah, Saudi Arabia

2Department of Medical Biochemistry, Sohag Faculty of Medicine, Sohag University, Egypt

3Department of Anatomy, Sohag Faculty of Medicine, Sohag University, Egypt

4Department of Anatomy, Al-Rayyan Medical College, Al-Madinah Al-Munawwarah, Saudi Arabia

5Department of Pathology, Sohag Faculty of Medicine, Sohag University, Egypt

6Department of Pathology, Al-Ghad Faculty of Health Sciences, Al-Madinah Al-Munawwarah, Saudi Arabia

7Department of Pediatrics, Tanta University Faculty of Medicine, Tanta, Gharbia, Egypt

8Department of Pediatrics, Aswan Faculty of Medicine, Aswan University, Egypt

9Department of Medical Biochemistry, Tanta Faculty of Medicine, Tanta University, Cairo, Egypt

100Department of Medicine, Taibah Faculty of Medicine, Taibah University, Al-Madinah Al-Munawwarah, Saudi Arabia

111Department of Family and Community Medicine, Faculty of Medicine, Sohag University, Egypt

122Division of cell biology, Ain Shams Faculty of Medicine, Egypt

133Department of anatomy and Embryology, Faculty of Medicine, Rabigh branch, King Abdul-Aziz University, Saudi Arabia

144Department of anatomy and Embryology, Faculty of Medicine, New Damietta, Al-Azhar University, Egypt

155Department of Ear, Nose and Throat, Taibah Faculty of Medicine, Taibah University, Al-Madinah Al-Munawwarah, Saudi Arabia

Pub. Date: March 21, 2018

Cite this paper:
Salah Mohamed El Sayed, Hussam H. Baghdadi, Nassar Ayoub Abdellatif Omar, Amal Nor Edeen Ahmad Allithy, Nahed Mohammed Hablas, Ahmed Ragab Fakhreldin, Reham Abelsalam Mariah, Momen El-Shazley, Mongi Ayat, Sayed Mostafa, Mostafa Abu-el Naga and Mohamed Abdel-Halim. The Antioxidant Glycolysis Inhibitor (Citric Acid) Induces a Dose-dependent Caspase-mediated Apoptosis and Necrosis in Glioma Cells. Journal of Cancer Research and Treatment. 2018; 6(1):18-24. doi: 10.12691/jcrt-6-1-4

Abstract

Background: Glioma tumors are still a big challenge being incurable with current chemotherapy and radiotherapy treatments. Surgical treatment of glioma needs adjuvant effective targeting therapy for better glioma cell treatment. Citrate is a well-known antioxidant organic acid abundant in citrus fruits and is an inhibitor of glycolysis through targeting the glycolytic enzyme phosphofructokinase, one of the key enzymes of glycolysis. Citrate is a natural product that is formed inside mitochondria during Krebs cycle to the extent that Krebs cycle is often referred to as citric acid cycle. It was reported that glioma cells are driven by glycolysis where glioma cells upregulates the expression of glycolysis genes and enzymes. Objectives: This aim is to investigate effect of citrate on glioma cells viability, morphology and moge of glioma-induced cell death. Methodology: In this study, citrate-induced glioma cell death was investigated using MTT assay, western blot analysis and flowcytometric evaluation was done to C6 glioma cells. Results: Citrate induced a potent anti-glioma effect by significantly decreasing viability of C6 glioma cells in a dose-dependent manner. Flow cytometric analysis revealed that at 5 mM, citrate induced a caspase-dependent apoptotic glioma cell death. Higher doses of citrate (9 mM) induced necro-apoptotic glioma cell death. Conclusion: citrate may be a promising therapeutic treatment for glioma and glioblastoma. Citrate-rich fruits are strongly recommended as a nutritional treatment for glioma patients.

Keywords:
citrate phosphofructokinase Glioblastoma apoptosis

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

Figures

Figure of 4

References:

[1]  Piao Y, Lu L, de Groot J. AMPA receptors promote perivascular glioma invasion via beta1 integrin-dependent adhesion to the extracellular matrix. Neuro Oncol. 2009 Jun; 11(3): 260-73.Epub 2008 Oct 28.
 
[2]  Nissler K, Petermann H, Wenz I, Brox D. Fructose 2,6-bisphosphate metabolism in Ehrlich ascites tumour cells. J Cancer Res Clin Oncol. 1995; 121(12): 739-45.
 
[3]  Rangel-Lopez E, Sanchez-Perez Y2, Soto-Reyes E3, Garcia-Cuellar CM2, Cacho-Diaz B2, Santamaria A. Role of epigenetics and oxidative stress in gliomagenesis. CNS Neurol Disord Drug Targets. 2018 Jan 10.
 
[4]  Kruspig B, Nilchian A, Orrenius S, Zhivotovsky B, Gogvadze V. Citrate kills tumor cells through activation of apical caspases. Cell Mol Life Sci. 2012 Dec; 69(24): 4229-37. Epub 2012 Oct 10.
 
[5]  Marie SK Shinjo SM. Metabolism and Brain Cancer. Clinics (Sao Paulo). 2011 Jun; 66(Suppl 1): 33-43.
 
[6]  Minchenko DO, Novik YE, Maslak HS, Tiazhka OV, Minchenko OH. EXPRESSION OF PFKFB, HK2, NAMPT, TSPAN13 AND HSPB8 GENES IN PEDIATRIC GLIOMA]. Lik Sprava. 2015 Oct-Dec; (7-8): 43-8.
 
[7]  Lu Y, Zhang X, Zhang H, Lan J, Huang G, Varin E, Lincet H, Poulain L, Icard P. Citrate induces apoptotic cell death: a promising way to treat gastric carcinoma? Anticancer Res. 2011 Mar; 31(3):797-805.
 
[8]  Guo X, Zhang X, Wang T, Xian S, Lu Y. 3-Bromopyruvate and sodium citrate induce apoptosis in human gastric cancer cell line MGC-803 by inhibiting glycolysis and promoting mitochondria-regulated apoptosis pathway. Biochem Biophys Res Commun. 2016 Jun 17; 475(1): 37-43.
 
[9]  Wang TA, Zhang XD, Guo XY, Xian SL, Lu YF. 3-bromopyruvate and sodium citrate target glycolysis, suppress survivin, and induce mitochondrial-mediated apoptosis in gastric cancer cells and inhibit gastric orthotopic transplantation tumor growth. Oncol Rep. 2016 Mar;35(3):1287-96
 
[10]  Kurhanewicz J, Dahiya R, Macdonald JM, Chang LH, James TL, Narayan P. Citrate alterations in primary and metastatic human prostatic adenocarcinomas: 1H magnetic resonance spectroscopy and biochemical study. Magn Reson Med. 1993 Feb; 29(2): 149-57.
 
[11]  Costello LC, Franklin RB, Narayan P. Citrate in the diagnosis of prostate cancer. Prostate. 1999 Feb 15; 38(3): 237-45.
 
[12]  Kline EE, Treat EG, Averna TA, Davis MS, Smith AY, Sillerud LO. Citrate concentrations in human seminal fluid and expressed prostatic fluid determined via 1H nuclear magnetic resonance spectroscopy outperform prostate specific antigen in prostate cancer detection. J Urol. 2006 Nov; 176(5): 2274-9.
 
[13]  Seymour ZA, Panigrahy A, Finlay JL, Nelson MD Jr, Blüml S. Citrate in pediatric CNS tumors? AJNR Am J Neuroradiol. 2008 May; 29(5): 1006-11.
 
[14]  Roelcke U, Leenders KL, von Ammon K, Radü EW, Vontobel P, Günther I, Psylla M. Brain tumor iron uptake measured with positron emission tomography and 52Fe-citrate. J Neurooncol. 1996 Aug; 29(2): 157-65.
 
[15]  Yousefi S, Owens JW, Cesario TC. Citrate shows specific, dose-dependent lympholytic activeity in neoplastic cell lines. Leuk Lymphoma. 2004 Aug; 45(8): 1657-65.
 
[16]  Islam MR, Samadi AR, Ahmed SM, Bardhan PK, Ali A. Oral rehydration therapy: efficacy of sodium citrate equals to sodium bicarbonate for correction of acidosis in diarrhoea. Gut. 1984 Aug; 25(8): 900-4.
 
[17]  Nagoba BS, Punpale AS, Ayachit R, Gandhi RC, Wadher BJ. Citric acid treatment of postoperative wound in an operated case of synovial sarcoma of the knee. Int Wound J. 2011 Aug; 8(4): 425-7. Epub 2011 May 12.
 
[18]  Salama N, Kishimoto T, Kanayama HO, Kagawa S. The mobile phone decreases fructose but not citrate in rabbit semen: a longitudinal study. Syst Biol Reprod Med. 2009 Dec; 55(5-6): 181-7.
 
[19]  Marín-Hernández A1, Rodríguez-Enríquez S, Vital-González PA, Flores-Rodríguez FL, Macías-Silva M, Sosa-Garrocho M, Moreno-Sánchez R. Determining and understanding the control of glycolysis in fast-growth tumor cells. Flux control by an over-expressed but strongly product-inhibited hexokinase. FEBS J. 2006 May; 273(9): 1975-88.
 
[20]  WARBURG O. On the origin of cancer cells. Science. 1956 Feb 24; 123(3191): 309-14.
 
[21]  Li XB, Gu JD, Zhou QH. Review of aerobic glycolysis and its key enzymes - new targets for lung cancer therapy. Thorac Cancer. 2015 Jan; 6(1): 17-24.
 
[22]  Grobben B, De Deyn PP, Slegers H. Rat C6 glioma as experimental model system for the study of glioblastoma growth and invasion. Cell Tissue Res. 2002 Dec; 310(3): 257-70.
 
[23]  Dominguez JE, Graham JF, Cummins CJ, Loreck DJ, Galarraga J, Van der Feen J, DeLaPaz R, Smith BH. Enzymes of glucose metabolism in cultured human gliomas: neoplasia is accompanied by altered hexokinase, phosphofructokinase, and glucose-6-phosphate dehydrogenase levels. Metab Brain Dis. 1987 Mar; 2(1): 17-30.
 
[24]  Gomez LS, Zancan P, Marcondes MC, Ramos-Santos L, Meyer-Fernandes JR, Sola-Penna M, Da Silva D. Resveratrol decreases breast cancer cell viability and glucose metabolism by inhibiting 6-phosphofructo-1-kinase. Biochimie. 2013 Jun; 95(6): 1336-43.
 
[25]  Marín-Hernández A, Rodríguez-Enríquez S, Vital-González PA, Flores-Rodríguez FL, Macías-Silva M, Sosa-Garrocho M, Moreno-Sánchez R. Determining and understanding the control of glycolysis in fast-growth tumor cells. Flux control by an over-expressed but strongly product-inhibited hexokinase. FEBS J. 2006 May; 273(9): 1975-88.
 
[26]  Dastidar SG, Sharma SK. Activities of glycolytic enzymes in rapidly proliferating and differentiated C6 glioma cells. Exp Cell Biol. 1989; 57(3): 159-64.
 
[27]  Kessler R, Bleichert F, Warnke JP, Eschrich K. 6-Phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB3) is up-regulated in high-grade astrocytomas. J Neurooncol. 2008 Feb; 86(3): 257-64.
 
[28]  Ismail R, Ul Hussain M. The up regulation of phosphofructokinase1 (PFK1) protein during chemically induced hypoxia is mediated by the hypoxia-responsive internal ribosome entry site (IRES) element, present in its 5'untranslated region. Biochimie. 2017 Aug; 139: 38-45
 
[29]  Sanzey M, Abdul Rahim SA, Oudin A, Dirkse A, Kaoma T, Vallar L, Herold-Mende C, Bjerkvig R, Golebiewska A, Niclou SP. Comprehensive analysis of glycolytic enzymes as therapeutic targets in the treatment of glioblastoma. PLoS One. 2015 May 1; 10(5): e0123544.
 
[30]  Auer RN, Del Maestro RF, Anderson R. A simple and reproducible experimental in vivo glioma model. Can J Neurol Sci. 1981 Nov; 8(4): 325-31.
 
[31]  Halabe Bucay A. Hypothesis proved...citric acid (citrate) does improve cancer: a case of a patient suffering from medullary thyroid cancer. Med Hypotheses. 2009 Aug; 73(2): 271.
 
[32]  Zhang, X., Varin, E., Allouche, S., Lu, Y., Poulain, L., Icard, P., 2009. Effect of citrate on malignant pleural mesothelioma cells: a synergistic effect with cisplatin. Anticancer Res. 29, 1249-1254.
 
[33]  Guo X, Zhang X, Wang T, Xian S, Lu Y. 3-Bromopyruvate and sodium citrate induce apoptosis in human gastric cancer cell line MGC-803 by inhibiting glycolysis and promoting mitochondria-regulated apoptosis pathway. Biochem Biophys Res Commun. 2016 Jun 17; 475(1): 37-43.
 
[34]  Hertz L, Peng L, Dienel GA. Energy metabolism in astrocytes: high rate of oxidative metabolism and spatiotemporal dependence on glycolysis/glycogenolysis. J Cereb Blood Flow Metab. 2007 Feb; 27(2): 219-49.
 
[35]  Szutowicz A, Kabata J, Bielarczyk H. The contribution of citrate to the synthesis of acetyl units in synaptosomes of developing rat brain. J Neurochem. 1982 May; 38(5): 1196-204.
 
[36]  Oudard S, Boitier E, Miccoli L, Rousset S, Dutrillaux B, Poupon MF. Gliomas are driven by glycolysis: putative roles of hexokinase, oxidative phosphorylation and mitochondrial ultrastructure. Anticancer Res. 1997 May-Jun; 17(3C): 1903-11.
 
[37]  Wolf A, Agnihotri S, Guha A. Targeting metabolic remodeling in glioblastoma multiforme. Oncotarget. 2010 Nov; 1(7): 552-62.