Copper Enhances Antitumor Activity of Water Soluble Garlic Extract Components on a Human Hepatoma Cell Line
Cancer Research Journal
Volume 4, Issue 5, September 2016, Pages: 73-83
Received: Aug. 3, 2016;
Accepted: Aug. 31, 2016;
Published: Sep. 21, 2016
Views 3076 Downloads 117
Angelo De Martino, Department of Biology, University of Rome “Tor Vergata”, Rome, Italy
Hanan Mahmoud Abu-Zeid, Botany Department, Faculty of Science, Alexandria University, Alexandria, Egypt
Piera Torricelli, Department SPES, University of Molise, Campobasso, Italy
Anna Shevchenko, Department of Science, People’s Friendship University of Russia, Moscow, Russia
Alberto Siciliano, Department of Biology, University of Rome “Tor Vergata”, Rome, Italy
Simone Beninati, Department of Biology, University of Rome “Tor Vergata”, Rome, Italy
Water garlic extract combined with Copper (GE-Cu) shows specific anti-proliferative and antitumoral activity in human hepatocarcinoma (HepG2) cancer cell line, while it is not active on differentiated or primary isolated cells, normal human dermal fibroblasts (HDF) and human epidermal keratinocytes (HEK). Since other transition metals do not affect cell viability, the observed antineoplastic property, was found specific for copper (Cu), and atomic absorption analyses demonstrate an accumulation of this metal inside the cell nuclei. GE-Cu treatment of HepG2a cancer cells, induces cell differentiation, growth arrest and programmed cell death. Furthermore, GE-Cu produces a continuous and prolonged flux of oxidative radicals, which down-regulates nuclear antioxidant defenses and repair systems, thus generating direct double-strand breaks to DNA. Indeed, it was observed that in GE-Cu-treated HepG2 cancer cells, histone deacetylase (HDAC) is inhibited, and histone H2AX is early phospho-activated. DNA double strand break downstream, likely responsible for the observed cell death in HepG2 cell, was evidenced by an early over expression of p53 and p21. JNK/c-Jun and p-38 phosphorylative cascade are activated as response to oxidative stress also. The resulting apoptosis appears to be caspases-independent because, under our experimental conditions, nuclear translocation of the apoptosis inducing factor (AIF) is operative. Our data suggest that GE-Cu possess an active specific antitumor capability, which could provide valuable new tools in cancer prevention and in supporting traditional chemotherapy.
Angelo De Martino,
Hanan Mahmoud Abu-Zeid,
Copper Enhances Antitumor Activity of Water Soluble Garlic Extract Components on a Human Hepatoma Cell Line, Cancer Research Journal.
Vol. 4, No. 5,
2016, pp. 73-83.
De Martino A, Torricelli P, Hanan Mahmoud Abu-Zeid, Shevchenko A, Siciliano A, Beninati S, Synergistic Anticancer Potential of Water Garlic Extract and Copper in a Human Hepatocarcinoma Cell Line. Cancer Research Journal 2016; 4 (2): 28-31.
Berthold HK, Sudhop T, von Bergmann K. Effect of a garlic oil preparation on serum lipoproteins and cholesterol metabolism: a randomized controlled trial. JAMA. 1998; 279: 1900-1902.
Amagase H, Petesch BL, Matsuura H, Kasuga S, Itakura Y. Intake of garlic and its bioactive components. J Nutr. 2001; 131: 955S-62S.
Bird SM, Ge H, Uden PC, Tyson JF, Block E, Denoyer E. High-performance liquid chromatography of selenoamino acids and organo selenium compounds. Speciation by inductively coupled plasma mass spectrometry. J Chromatogr A. 1997; 789: 349-359.
Dumont E, Ogra Y, Vanhaecke F, Suzuki KT, Cornelis R. Liquid chromatography-mass spectrometry (LC-MS): a powerful combination for selenium speciation in garlic (Allium sativum). Anal Bioanal Chem. 2006; 384: 1196-1206.
Shirin H, Pinto JT, Kawabata Y, Soh JW, Delohery T, Moss SF, Murty V, Rivlin RS, Holt PR, Weinstein IB. Antiproliferative effects of S-allylmercaptocysteine on colon cancer cells when tested alone or in combination with sulindac sulfide. Cancer Res. 2001; 61: 725-731.
Xiao D, Pinto JT, Gundersen GG, Weinstein IB. Effects of a series of organosulfur compounds on mitotic arrest and induction of apoptosis in colon cancer cells. Mol Cancer Ther. 2005; 4: 1388-1398.
Ho JW, Jie M. Pharmacological activity of cardiovascular agents from herbal medicine. Cardiovasc Hematol Agents Med Chem. 2007; 5: 273-277.
Ghayur MN, Khan H, Gilani AH. Antispasmodic, bronchodilator and vasodilator activities of (+)-catechin, a naturally occurring flavonoid. Arch Pharm Res. 2007; 30: 970-975.
Comalada M, Ballester I, Bailon E, Sierra S, Xaus J, Galvez J, de Medina FS, Zarzuelo A. Inhibition of pro-inflammatory markers in primary bone marrow-derived mouse macrophages by naturally occurring flavonoids: analysis of the structure-activity relationship. Biochem Pharmacol. 2006; 16; 72: 1010-1021.
Filipe P, Lanca V, Silva JN, Morliere P, Santus R, Fernandes A. Flavonoids and urate antioxidant interplay in plasma oxidative stress. Mol Cell Biochem. 2001; 221: 79-87.
Brown JE, Khodr H, Hider RC, Rice-Evans CA. Structural dependence of flavonoid interactions with Cu2+ ions: implications for their antioxidant properties. Biochem J. 1998; 330 (Pt 3): 1173-1178.
Pereira RM, Andrades NE, Paulino N, Sawaya AC, Eberlin MN, Marcucci MC, Favero GM, Novak EM, Bydlowski SP. Synthesis and characterization of a metal complex containing naringin and Cu, and its antioxidant, antimicrobial, antiinflammatory and tumor cell cytotoxicity. Molecules. 2007; 12: 1352-1366.
Hadi SM, Bhat SH, Azmi AS, Hanif S, Shamim U, Ullah MF. Oxidative breakage of cellular DNA by plant polyphenols: a putative mechanism for anticancer properties. Semin Cancer Biol. 2007; 17: 370-376.
Valko M, Rhodes CJ, Moncol J, Izakovic M, Mazur M. Free radicals, metals and antioxidants in oxidative stress-induced cancer. Chem Biol Interact. 2006; 160: 1-40.
Yamazaki T, Yamazaki A, Onuki H, Hibino Y, Yokote Y, Sakagami H, Nakajima H, Shimada J. Effect of saliva, epigallocatechin gallate and hypoxia on Cu-induced oxidation and cytotoxicity. In Vivo. 2007; 21: 603-607.
Yu HN, Shen SR, Yin JJ. Effects of metal ions, catechins, and their interactions on prostate cancer. Crit Rev Food Sci Nutr. 2007; 47: 711-719.
Ogita A, Fujita K, Taniguchi M, Tanaka T. Dependence of synergistic fungicidal activity of Cu2+ and allicin, an allyl sulfur compound from garlic, on selective accumulation of the ion in the plasma membrane fraction via allicin-mediated phospholipid peroxidation. Planta Med. 2006; 72: 875-880.
Ogita A, Hirooka K, Yamamoto Y, Tsutsui N, Fujita K, Taniguchi M, Tanaka T. Synergistic fungicidal activity of Cu (2+) and allicin, an allyl sulfur compound from garlic, and its relation to the role of alkyl hydroperoxide reductase 1 as a cell surface defense in Saccharomyces cerevisiae. Toxicology. 2005; 215: 205-213.
Uauy R, Olivares M, Gonzalez M. Essentiality of copper in humans. Am J Clin Nutr. 1998; 67: 952S-959S.
Halliwell B, Gutteridge JM. Role of free radicals and catalytic metal ions in human disease: an overview. Methods Enzymol. 1990; 186: 1-85.
Pellegrini G, Ranno R, Stracuzzi G, Bondanza S, Guerra L, Zambruno G, Micali G, De Luca M. The control of epidermal stem cells (holoclones) in the treatment of massive full-thickness burns with autologous keratinocytes cultured on fibrin. Transplantation. 1999; 68: 868-879.
Korn JH. Fibroblast prostaglandin E2 synthesis. Persistence of an abnormal phenotype after short-term exposure to mononuclear cell products. J Clin Invest. 1983; 71: 1240-1246.
Alisi A, Leoni S, Piacentani A, Conti Devirgiliis L. Retinoic acid modulates the cell-cycle in fetal rat hepatocytes and HepG2 cells by regulating cyclin-cdk activities. Liver Int. 2003; 23: 179-186.
Ciriolo MR, Aquilano K, De Martino A, Carri MT, Rotilio G. Differential role of superoxide and glutathione in S-nitrosoglutathione-mediated apoptosis: a rationale for mild forms of familial amyotrophic lateral sclerosis associated with less active Cu, Zn superoxide dismutase mutants. J Neurochem. 2001; 77: 1433-1443.
Filomeni G, Rotilio G, Ciriolo MR. Glutathione disulfide induces apoptosis in U937 cells by a redox-mediated p38 MAP kinase pathway. FASEB J. 2003; 17: 64-66.
Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 1991; 139: 271-279.
Vigilanza P, Aquilano K, Rotilio G, Ciriolo MR. Transient cytoskeletal alterations after SOD1 depletion in neuroblastoma cells. Cell Mol Life Sci. 2008; 65: 991-1004.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193: 265-275.
McCormick DL, Johnson WD, Kozub NM, Rao KV, Lubet RA, Steele VE, Bosland MC. Chemoprevention of rat prostate carcinogenesis by dietary 16alpha-fluoro-5-androsten-17-one (fluasterone), a minimally androgenic analog of dehydroepiandrosterone. Carcinogenesis. 2007; 28: 398-403.
Saiko P, Szakmary A, Jaeger W, Szekeres T. Resveratrol and its analogs: defense against cancer, coronary disease and neurodegenerative maladies or just a fad? Mutat Res. 2008; 658: 68-94.
Seren S, Lieberman R, Bayraktar UD, Heath E, Sahin K, Andic F, Kucuk O. Lycopene in cancer prevention and treatment. Am J Ther. 2008; 15: 66-81.
Yamanoshita O, Ichihara S, Hama H, Ichihara G, Chiba M, Kamijima M, Takeda I, Nakajima T. Chemopreventive effect of selenium-enriched Japanese radish sprout against breast cancer induced by 7, 12-dimethylbenz[a]anthracene in rats. Tohoku J Exp Med. 2007; 212: 191-198.
Jadhav U, Ezhilarasan R, Vaughn SF, Berhow MA, Mohanam S. Dietary isothiocyanate iberin inhibits growth and induces apoptosis in human glioblastoma cells. J Pharmacol Sci. 2007; 103: 247-251.
Letavayova L, Vlckova V, Brozmanova J. Selenium: from cancer prevention to DNA damage. Toxicology. 2006; 227: 1-14.
Filomeni G, Aquilano K, Rotilio G, Ciriolo MR. Reactive oxygen species-dependent c-Jun NH2-terminal kinase/c-Jun signaling cascade mediates neuroblastoma cell death induced by diallyl disulfide. Cancer Res. 2003; 63: 5940-5949.
Belloir C, Singh V, Daurat C, Siess MH, Le Bon AM. Protective effects of garlic sulfur compounds against DNA damage induced by direct- and indirect-acting genotoxic agents in HepG2 cells. Food Chem Toxicol. 2006; 44: 827-834.
Macho A, Calzado MA, Munoz-Blanco J, Gomez-Diaz C, Gajate C, Mollinedo F, Navas P, Munoz E. Selective induction of apoptosis by capsaicin in transformed cells: the role of reactive oxygen species and calcium. Cell Death Differ. 1999; 6: 155-165.
Rudolf E, Rudolf K, Cervinka M. Selenium activates p53 and p38 pathways and induces caspase-independent cell death in cervical cancer cells. Cell Biol Toxicol. 2008; 24: 123-141.
Jiang C, Hu H, Malewicz B, Wang Z, Lu J. Selenite-induced p53 Ser-15 phosphorylation and caspase-mediated apoptosis in LNCaP human prostate cancer cells. Mol Cancer Ther. 2004; 3: 877-884.
Milaeva, E. R. Metal-based antioxidants--potential therapeutic candidates for prevention the oxidative stress-related carcinogenesis: mini-review. Curr Top Med Chem. 2011, 11 (21), 2703-2713.
Pierpoint, W. S.; Harrison, B. D. Cu-Dependent and Iron-Dependent Inactivations of Cucumber Mosaic Virus by Polyphenols. J Gen Microbiol. 1963, 32: 429-440.
Javadi, F.; Eghtesadi, S.; Ahmadzadeh, A.; Aryaeian, N.; Zabihiyeganeh, M.; Foroushani, A. R.; Jazayeri, S. The effect of quercetin on plasma oxidative status, C-reactive protein and blood pressure in women with rheumatoid arthritis. Int J Prev Med. 2014, 5 (3), 293-301.
Noguchi, N.; Niki, E. Phenolic antioxidants: a rationale for design and evaluation of novel antioxidant drug for atherosclerosis. Free Radic Biol Med. 2000, 28 (10), 1538-1546.
Sarma, R. J.; Otto, S.; Nitschke, J. R. Disulfides, imines, and metal coordination within a single system: interplay between three dynamic equilibria. Chemistry. 2007, 13 (34), 9542-9546.