Combination of Paracetamol and the Glutathione Depleting Agent Buthionine Sulfoximine Show Differential Effect on Liver Cancer Cells and Normal Hepatocytes — Oak Academic Publishing
Research ArticleOpen AccessGoogle Scholar indexed
Combination of Paracetamol and the Glutathione Depleting Agent Buthionine Sulfoximine Show Differential Effect on Liver Cancer Cells and Normal Hepatocytes
National Cancer Institute, Cairo University, Cairo, Egypt
,
Department of Pharmacology & Toxicology, Faculty of Pharmacy, Cairo University, Cairo, Egypt
,
Department of Pharmacology & Toxicology, Faculty of Pharmacy, Cairo University, Cairo, Egypt
,
Department of Pathology, National Cancer Institute, Cairo University, Cairo, Egypt
,
Sharjah Institute for Medical Research and College of Pharmacy, University of Sharjah, Sharjah, UAE
1 National Cancer Institute, Cairo University, Cairo, Egypt
2 Department of Pharmacology & Toxicology, Faculty of Pharmacy, Cairo University, Cairo, Egypt
3 Department of Pharmacology & Toxicology, Faculty of Pharmacy, Cairo University, Cairo, Egypt
4 Department of Pathology, National Cancer Institute, Cairo University, Cairo, Egypt
5 Sharjah Institute for Medical Research and College of Pharmacy, University of Sharjah, Sharjah, UAE
Background : Paracetamol exerts toxic effects on liver cells through its metabolism into N-acetyl-p-benzoquinone imine (NAPQI), which is detoxified by conjugation with cellular glutathione (GSH). Once GSH is depleted, NAPQI stimulates a range of oxidative reactions that result in cell necrosis. The aim of the present investigation is to find a new strategy that would selectively protect normal hepatic tissues and sensitize liver cancer cells to the toxic effects of paracetamol or its metabolite. This may lead to the development of a targeted therapy for liver cancer. Methods : The anti-proliferative effects of paracetamol and buthionine sulfoximine BSO (a glutathione depleting agent) alone and in combination on the liver cancer cells HepG2 and normal rat hepatocytes were investigated by sulphorhodamine-B assay. Effects on cell cycle regulation and induction of apoptosis were tested by flow cytometry. The level of prostaglandin expression was measured by ELISA. Results : The present study showed that both agents alone or in combination have anti-proliferative effects on both cell types. Surprisingly, BSO showed a cytoprotective effects on normal hepatocytes treated with high concentrations (1.75 and 2 mM) of paracetamol. This was confirmed by cell cycle analysis that recorded decreased fraction of sub-G1 cells indicating reduction of apoptosis in normal hepatocytes. Analysis of prostaglandin E2 revealed differential effects of paracetamol on normal and liver cancer cells. A significant increase in PGE2 level over the control was observed in normal hepatocytes whereas a significant decrease was seen in HepG2 cells after treatment with paracetamol. Conclusion : These results indicate that combination of paracetamol/BSO has differential effects on liver cancer cells and normal hepatocytes, which opens the avenue for a new effective and selective combination for management of liver cancer.
Fong, Y., Dupuy, D.E., Feng, M. and Abou-Alfa, G. (2015) Cancer of the Liver. In: De Vita, V.T., Hellman, S., Rosenberg, S.A., Eds., Cancer: Principles and Practice of Oncology, 10th Edition, Lippincott Williams & Wilkins, Philadelphia, 123.
Manov, I., Hirsh, M. and Iancu, T.C. (2004) N-Acetylcysteine Does Not Protect HepG2 Cells against Acetaminophen-Induced Apoptosis. Basic & Clinical Pharmacology & Toxicology, 94, 213-225. http://dx.doi.org/10.1111/j.1742-7843.2004.pto940504.x
Dai, Y. and Cederbaum, A.I. (1995) Cytotoxicity of Acetaminophen in Human Cytochrome P4502E1-Transfected HepG2 Cells. Journal of Pharmacology and Experimental Therapeutics, 273, 1497-1505.
Nelson, S.D. (1995) Mechanisms of the Formation and Disposition of Reactive Metabolites That Can Cause Acute Liver Injure. Drug Metabolism Reviews, 27, 147-177. http://dx.doi.org/10.3109/03602539509029821
Tonge, R.P., Kelly, E.J., Bruschi, S.A., Kalhorn, T., Eaton, D.L., Nebert, D. and Nelson, S.D. (1998) Role of CYP1A2 in the Hepatotoxicity of Acetaminophen: Investigation Using Cyp1a2 Null Mice. Toxicology and Applied Pharmacology, 153, 102-108. http://dx.doi.org/10.1006/taap.1998.8543
Manyike, P.T., Kharasch, E.D., Kalhorn, T.F. and Slattery, J.T. (2000) Contribution of CYP2E1 and CYP3A to Acetaminophen Reactive Metabolite Formation. Clinical Pharmacology & Therapeutics, 67, 275-282. http://dx.doi.org/10.1067/mcp.2000.104736
Liang, Y.L., Zhang, Z.H., Liu, X.J., Liu, X.Q., Tao, L., Zhang, Y.F., et al. (2012) Melatonin Protects against Apoptosis-Inducing Factor (AIF) Dependent Cell Death during Acetaminophen-Induced Acute Liver Failure. PLoS ONE, 7, e51911. http://dx.doi.org/10.1371/journal.pone.0051911
Zhao, X., Cong, X., Zheng, L., Xu, L., Yin, L. and Peng, J. (2012) Dioscin, a Natural Steroid Saponin, Shows Remarkable Protective Effect against Acetaminophen-Induced Liver Damage in Vitro and in Vivo. Toxicology Letters, 214, 69-80. http://dx.doi.org/10.1016/j.toxlet.2012.08.005
Biaglow, J.E., Clark, E.P., Epp, E.R., Morse-Guardio, M., Varnes, M.E. and Mitchell, J.B. (1983) Non-Protein Thiols and the Radiation Response of A549 Human Lung Carcinoma Cells. International Journal of Radiation Biology, 44, 489-495.
Mitchell, J.B., Morstyn, G., Russo, A. and Carney, D.N. (1985) In Vitro Radiobiology of Human Lung Cancer. Cancer Treatment Symposia, 2, 3-10.
Rogers, L.K., Moorthy, B. and Smith, C.V. (1997) Acetaminophen Binds to Mouse Hepatic and Renal DNA at Human Therapeutic Doses. Chemical Research in Toxicology, 10, 470-476. http://dx.doi.org/10.1021/tx960159i
Kaplowitz, N. (2004) Acetaminophen Hepatoxicity: What Do We Know, What Don’t We Know, and What Do We Do Next? Hepatology, 40, 23-26. http://dx.doi.org/10.1002/hep.20312
Rouzer, C.A., Scott, W.A., Griffith, W., Hamill, A.L. and Cohn, Z.A. (1982) Arachidonic Acid Metabolism in Glutathione-Deficient Macrophages. Proceedings of the National Academy of Sciences of the United States of America, 79, 1621-1625. http://dx.doi.org/10.1073/pnas.79.5.1621
Anderson, C.P., Tsai, J.M., Meek, W.E., Liu, R.M., Tang, Y., Forman, H.J. and Reynolds, C.P. (1999) Depletion of Glutathione by Buthionine Sulfoxine Is Cytotoxic for Human Neuroblastoma Cell Lines via Apoptosis. Experimental Cell Research, 246, 183-192. http://dx.doi.org/10.1006/excr.1998.4303
Seglen, P.O. (1976) Preparation of Isolated Rat Liver Cells: The Enzymatic Preparation of Isolated Intact Parenchymal Cells from Rat Liver. Methods in Cell Biology, 13, 29-83. http://dx.doi.org/10.1016/S0091-679X(08)61797-5
El-Awady, R.A., Semreen, M.H., Saber-Ayad, M.M., Cyprian, F., Menon, V. and Al-Tel, T.H. (2016) Modulation of DNA Damage Response and Induction of Apoptosis Mediates Synergism between Doxorubicin and a New Imidazopyridine Derivative in Breast and Lung Cancer Cells. DNA Repair, 37, 1-11. http://dx.doi.org/10.1016/j.dnarep.2015.10.004
El-Awady, R.A., Saleh, E.M., Ezz, M. and Elsayed, A.M. (2011) Interaction of Celecoxib with Different Anti-Cancer Drugs Is Antagonistic in Breast but Not in Other Cancer Cells. Toxicology and Applied Pharmacology, 255, 271-286. http://dx.doi.org/10.1016/j.taap.2011.06.019
Sumida, A., Fukuen, S., Yamamoto, I., Matsuda, H., Naohara, M. and Azuma, J. (2000) Quantitative Analysis of Constitutive and Inducible SYPs mRNA Expression in the HepG2 Cell Line Using Reverse Transcription-Competitive PCR. Biochemical and Biophysical Research Communications, 267, 756-760. http://dx.doi.org/10.1006/bbrc.1999.2029
Fath, M.A., Ahmad, I.M., Smith, C.J., Spence, J. and Spitz, D.R. (2011) Enhancement of Carboplatin-Mediated Lung Cancer Cell Killing by Simultaneous Disruption of Glutathione and Thioredoxin Metabolism. Clinical Cancer Research, 17, 6206-6017. http://dx.doi.org/10.1158/1078-0432.CCR-11-0736
Schnelldorfer, T., Gansauge, S., Gansauge, F., Schlosser, S., Beger, H.G. and Nussler, A.K. (2000) Glutathione Depletion Causes Cell Growth Inhibition and Enhanced Apoptosis in Pancreatic Cancer Cells. Cancer, 89, 1440-1447. http://dx.doi.org/10.1002/1097-0142(20001001)89:7 3.0.CO;2-0
Shaw, J.P. and Chou, L.N. (1986) Elevation of Intracellular Glutathione Content Associated with Mitogenic Stimulation of Quiescent Fibroblasts. Journal of Cellular Physiology, 129, 193-198. http://dx.doi.org/10.1002/jcp.1041290210
Syng-Ai, C., Kumari, A.L. and Khar, A. (2004) Effect of Curcumin on Normal and Tumor Cells: Role of Glutathione and Bcl-2. Molecular Cancer Therapeutics, 3, 1101-1108.
Lee, F.Y.F., Allalunis-Turner, M.J. and Siemann, D.W. (1987) Depletion of Tumor versus Normal Tissue Glutathione by Buthionine Sulfoximine. British Journal of Cancer, 56, 33-38. http://dx.doi.org/10.1038/bjc.1987.148
Honn, K.V., Bockman, R.S. and Marnett, L.J. (1981) Prostaglandins and Cancer: A Review of Tumor Initiation through Tumor Metastasis. Prostaglandins, 21, 833-850. http://dx.doi.org/10.1016/0090-6980(81)90240-9
Honn, K.V. and Sloane, B.F. (1985) Prostaglandins in Tumor Cell Metastasis. In: Torisv, M. and Yoshida, T., Eds., Basic Mechanisms and Clinical Treatment of Tumor Metastasis, Academic Press, New York, 311-334. http://dx.doi.org/10.1016/B978-0-12-695680-1.50024-7
Meerpohl, H.G., Bauknecht, T., Tritschler, V. and Lang, H. (1981) The Influence of Prostaglandin E2 on Macrophage Mediated Cytotoxicity. In: Forster, W., Ed., International Workshop: Heterogeneity of Mononuclear Phagocytes, Academic Press, New York, 428-461.
Phipps, R.P., Stein, S.H. and Roper, R.L. (1991) A New View of Prostaglandin E Regulation of the Immune Response. Immunology Today, 12, 349-352. http://dx.doi.org/10.1016/0167-5699(91)90064-Z
Furstenberger, G., Gross, M. and Marks, F. (1984) Involvement of Prostaglandins in the Process of Skin Tumor Promotion. In: Thaler-Dao, H., Crastes de Paulet, A. and Paoletti, R., Eds., Icosanoids and Cancer, Raven Press, New York, 91-100.
Furstenberger, G., Gross, M. and Marks, F. (1984) On the Role of Prostaglandins in the Induction of Epidermal Proliferation, Hyperplasia and Tumor Promotion in Mouse Skin. In: Powles, T.J., Bockman, R.S., Honn, K.V. and Ramwell, P.W., Eds., Prostaglandins and Related Lipids, Vol. 2, Alan R Liss, New York, 239-251.
Hubbord, W.C., Alley, M.C., McLemore, T.L. and Boyd, M.R. (1988) Profiles of Prostaglandin Biosynthesis in Sixteen Established Cell Lines Derived from Human Lung, Colon, Prostate, and Ovarian Tumors. Cancer Research, 48, 4770-4775.
Green, K., Drvota, V. and Vesterqvist, O. (1989) Pronounced Reduction of in Vivo Prostacyclin Synthesis in Humans by Acetaminophen (Paracetamol). Prostaglandins, 37, 311-315. http://dx.doi.org/10.1016/0090-6980(89)90001-4
Lee, Y.S., Kim, H., Brahim, J.S., Rowan, J., Lee, G. and Donne, R.A. (2007) Acetaminophen Selectively Suppresses Peripheral Prostaglandin E2 Release and Increases COX2 Gene Expression in a Clinical Model of Acute Inflammation. Pain, 129, 279-286. http://dx.doi.org/10.1016/j.pain.2006.10.020
Crawley, B., Saito, O., Malkmus, S., Fitzsimmons, B., Hua, X.Y. and Yaksh, T.L. (2008) Acetaminophen Prevents Hyperalgesia in Central Pain Cascade. Neuroscience Letters, 442, 50-53. http://dx.doi.org/10.1016/j.neulet.2008.06.062
Kehoe, M.J., Cohen, S.M., Zarrinnia, K. and Cowan, A. (1996) The Effect of Acetaminophen, Ibuprofen and Misoprostol on Prostaglandin E2 Synthesis and the Degree and Rate of Orthodontic Tooth Movement. Angle Orthodontist, 66, 339-350.
Botting, R.M. (2000) Mechanism of Action of Acetaminophen: Is There a Cyclooxygenase 3? Clinical Infectious Diseases, 31, S202-S210. http://dx.doi.org/10.1086/317520
Kis, B., Snipes, J.A., Simandle, S.A. and Busija, D.W. (2005) Acetaminophen-Sensitive Prostaglandin Production in Rat Cerebral Endothelial Cells. American Journal of Physiology-Regulatory Integrative and Comparative Physiology, 288, R897-R902. http://dx.doi.org/10.1152/ajpregu.00613.2004
Hanel, A.M. and Lands, W.E.M. (1982) Modification of Anti-Inflammatory Drug Effectiveness by Ambient Lipid Peroxides. Biochemical Pharmacology, 31, 3307-3311. http://dx.doi.org/10.1016/0006-2952(82)90565-2
Hertz, F. and Cloarec, A. (1984) Pharmacology of Free Radicals, Recent Views on Their Relation to Inflammatory Mechanisms. Life Sciences, 34, 713-720. http://dx.doi.org/10.1016/0024-3205(84)90378-3
Graham, G.G., Davies, M.J., Day, R.O., Mohamudally, A. and Scott, K.F. (2013) The Modern Pharmacology of Paracetamol: Therapeutic Actions, Mechanism of Action, Metabolism, Toxicity and Recent Pharmacological Findings. Inflammopharmacology, 21, 201-232. http://dx.doi.org/10.1007/s10787-013-0172-x
Simmons, D.L., Wagner, D. and Westover, K. (2000) Nonsteroidal Anti-Inflammatory Drugs, Acetaminophen, Cyclooxygenase 2, and Fever. Clinical Infectious Diseases, 31, S211-S218. http://dx.doi.org/10.1086/317517