The aim of the present study was to compare the ability of the myricetin and quercetin to modulate the oxidative DNA damage induced by 2-amino-3, 8- dimethylimidazo [4,5-f] quinoxaline (8-MeIQx), 2-amino- 3, 4, 8- trimethylimidazo [4, 5-f]-quinoxaline (4,8-diMeIQx) and 2-amino-1-methyl-6-phenyl-imidazo [4,5-b] pyridine (PhIP), in human hepatoma cells. DNA damage (strand breaks and oxidized purines/pyrimidines) was evaluated by the alkaline single-cell gel electrophoresis or comet assay. None of the myricetin and quercetin concentrations tested protected against 8-MeIQx, 4, 8-diMeIQx and PhIP-induced DNA strand breaks. The oxidized pyrimidines induced by 4, 8-diMeIQx and PhIP were reduced by myricetin but not by quercetin. Quercetin reduced the oxidized purines induced by 8-MeIQx and PhIP, while myricetin also reduced the induced by 4, 8-diMeIQx. One feasible mechanism by which myricetin and quercetin exert their protective effect towards HCAs-induced oxidative DNA could be related in part to the reduction of human CYP1A1. Another mechanism claimed to be responsible for the protective effect of myricetin and quercetin is the induction of phase II metabolizing enzymes such as UDP-glucuronyltrasferase (UGT). The ethoxyresorufin O-deethylation (CYP1A1) activity was moderately inhibited by myricetin, while little effect was observed by quercetin. On the contrary, quercetin showed the greatest increase on UDP-glucuronyltransferase activity. However, these are not the only mechanisms by which myricetin and quercetin exert their protective effect, other mechanisms such as stimulation of the repair of carcinogen-induced DNA damage and or the free radical scavenging efficiency have been also implicated. In conclusion, our results clearly indicate that myricetin was more efficient than quercetin to prevent DNA damage (oxidized purines and pyrimidines) induced by the three HCAs evaluated. This protective effect depends on the chemical structure of flavonoid and the mutagen studied.
KeywordsMyricetinQuercetinComet AssayOxidative DNA DamageHeterocyclic Amines
World Cancer Research Fund, “Food, Nutrition and the Prevention of Cancer: A Global Perspective,” Nutrtion, Vol. 15, No. 6, 1997, pp. 523-526.
S. J. Duthie, A. R. Collins, G. C. Duthie and V. L. Dobson, “Quercetin and Myricetin Protect against Hydrogen Peroxide-Induced DNA Damage Strand Breaks and Oxidized Pyrimidines, in Human Lymphocytes,” Mutation Research, Vol. 393, No. 3, 1997, pp. 223-231.
M. Kampa, A. Nifli, G. Notas and E. Castanas, “Polyphenols and Cancer Cell Growth,” Reviews of Physiology, Biochemistry & Pharmacology, Vol. 159, No. 2, 2007, pp. 79-113. doi:10.1007/112_2006_0702
K. Min and S. E. Ebeler, “Flavonoid Effects on DNA Oxidation at Low Concentrations Relevant to Physiological Levels,” Food and Chemical Toxicology, Vol. 46, No. 1, 2008, pp. 96-104. doi:10.1016/j.fct.2007.07.002
M. Maggiolini, A. G. Recchia, D. Bonofiglio, S. Catalano, A. Vivacqua, A. Carpino, V. Rago, R. Rossi and S. Ando, “The Red Wine Phenolics Piceatannol and Myricetin Act as Agonists for Estrogen Receptor {Alpha} in Human Breast Cancer Cells,” Journal of Molecular Endocrinology, Vol. 35, No. 2, 2005, pp. 269-281. doi:10.1677/jme.1.01783
M. L. Neuhouser, “Dietary Flavonoids and Cancer Risk: Evidence From Human Population Studies,” Nutrition and Cancer, Vol. 50, No. 1, 2004, pp. 1-7. doi:10.1207/s15327914nc5001_1
T. Sugimura, “Overview of Carcinogenic Heterocyclic Amines,” Mutation Research, Vol. 376, No. 1, 1997, pp. 211-219. doi:10.1016/S0027-5107(97)00045-6
R. Sinha, “An Epidemiologic Approach to Studying Heterocyclic Amines,” Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, Vol. 506-507, 2002, pp. 197-204. doi:10.1016/S0027-5107(02)00166-5
J. Felton and M. Malfatti, “What Do Diet-Induced Changes in Phase I and II Enzymes Tell Us about Prevention from Exposure to Heterocyclic Amines?” Journal of Nutrition, Vol. 136, No. 10, 2006, pp. 2683-2684.
F. Toribio, R. Busquets, L. Puignou and M. T. Galceran, “Heterocyclic Amines in Griddled Beef Steak Analysed Using a Single Extract Clean-Up Procedure,” Food and Chemical Toxicology, Vol. 45, No. 4, 2007, pp. 667-675. doi:10.1016/j.fct.2006.10.016
A. Ristic, M. Cichna and G. Sontag, “Determination of Less Polar Heterocyclic Aromatic Amines in Standardised Beef Extracts and Cooked Meat Consumed in Austria by Liquid Chromatography and Fluorescence Detection,” Journal of Chromatography B, Vol. 802, No. 1, 2004, pp. 87-94. doi:10.1016/j.jchromb.2003.09.028
K. I. Skog, M. A. E. Johansson and M. I. J?gerstad, “Carcinogenic Heterocyclic Amines in Model Systems and Cooked Foods: A Review on Formation, Occurrence and Intake,” Food and Chemical Toxicology, Vol. 36, No. 9-10, 1998, pp. 879-896. doi:10.1016/S0278-6915(98)00061-1
International Agency for Research on Cancer, “Some Natural Occurring Substances: Food Items and Constituents Heterocyclic Amines and Mycotoxins: IARC Monographs on the Evaluation of Carcinogenic Risks to Humans by International Agency for Research on Cancer,” World Health Organization, Lyon, 1993, pp. 165-231.
S. Knasmuller, C. E. Schwab, S. J. Land, C. Y. Wang, R. Sanyal, M. Kundi, W. Parzefall and F. Darroudi, “Genotoxic Effects of Heterocyclic Aromatic Amines in Human Derived Hepatoma (HepG2) Cells,” Mutagenesis, Vol. 14, No. 6, 1999, pp. 533-540. doi:10.1093/mutage/14.6.533
J. Felton, M. Knize, L. M. Bennett, M. Malfatti, M. Col- vin and K. Kulp, “Impact of Environmental Exposures on the Mutagenicity/Carcinoge Nicity of Heterocyclic Ami- nes,” Toxicology, Vol. 198, No. 1-3, 2004, pp. 135-145. doi:10.1016/j.tox.2004.01.024
Y. He, M. D. Friesen, R. J. Ruch and H. A. J. Schut, “Indole-3-Carbinol as a Chemopreventive Agent in 2-Am- ino-1-Methyl- 6-Phenyl Imdazo [4,5-b] Pyridine (PhIP) Carcinogenesis: Inhibition of PhIP–DNA Adduct Formation, Acceleration of PhIP Metabolism, and Induction of Cytochrome P450 in Female F344 Rats,” Food and Che- mical Toxicology, Vol. 38, No. 1, 2000, pp. 15-23. doi:10.1016/S0278-6915(99)00117-9
R. H. Dashwood, “Modulation of Heterocyclic Amine-In- duced Mutagenicity and Carcinogenicity: An ‘A-to-Z’ Guide to Chemopreventive Agents, Promoters, and Tran- sgenic Models,” Mutation Research, Vol. 511, No. 2, 2002, pp. 89-112. doi:10.1016/S1383-5742(02)00005-4
A. R. Collins, “Assays for Oxidative Stress and Antioxidant Status: Applications to Research into the Biological Effectiveness of Polyphenols,” The American Journal of Clinical Nutrition, Vol. 81, No. 1, 2005, pp. 261-267.
V. Mersch-Sundermann, S. Knasmuller, X. Wu, F. Dar- roudi and F. Kassie, “Use of a Human-Derived Liver Cell Line for the Detection of Cytoprotective, Antigenotoxic and Cogenotoxic Agents,” Toxicology, Vol. 198, No. 1-3, 2004, pp. 329-340. doi:10.1016/j.tox.2004.02.009
M. A. Westerink and G. E. J. Schoonen, “Cytochrome P450 Enzyme Levels in HepG2 Cells and Cryopreserved Primary Human Hepatocytes and Their Induction in HepG2 Cells,” Toxicology in Vitro, Vol. 21, No. 8, 2007, pp. 1581-1591. doi:10.1016/j.tiv.2007.05.014
M. E. Delgado, A. I. Haza, N. Arranz, A. García and P. Morales, “Dietary Polyphenols Protect against N-Nitro- samines and Benzo(a) Pyrene-Induced DNA Damage (Str- and Breaks and Oxidized Purines/Pyrimidines) in HepG2 Human Hepatoma Cells,” European Journal of Nutrition, Vol. 47, No. 11, 2008, pp. 479-490. doi:10.1007/s00394-008-0751-6
P. L. Olive, D. Wlodek, R. E. Durand and J. P. Banáth, “Factors Influencing DNA Migration from Individual Cells Subjected to Gel Electrophoresis,” Experimental Cell Research, Vol. 198, No. 2, 1992, pp. 259-267. doi:10.1016/0014-4827(92)90378-L
A. I. Haza and P. Morales, “Effects of (+) Catechin and (-) Epicatechin on Heterocyclic Amines-Induced Oxidative DNA Damage,” Journal of Applied Toxicology, Vol. 31, No. 1, 2011, pp. 53-62. doi:10.1002/jat.1559
M. D. Burke and R. T. Mayer, “Differential Effects of Phenobarbitone and 3-Methyl Holanthrene Induction on the Hepatic Microsomal Metabolism and Cytochrome P-450-Binding of Phenoxazone and a Homologous Series of Its n-Alkyl Ethers (Alkoxyresorufins),” Chemico-Bio- logical Interactions, Vol. 45, No. 2, No. 2, 1983, pp. 243-258. doi:10.1016/0009-2797(83)90072-8
M. D. Burke, S. Thompson, C. R. Elcombe, J. Halpert, T. Haaparanta and R. T. Mayer, “Ethoxy-Penthoxy and Benzyloxyphexazones and Homologues: A Series of Sub- strates to Different Induced Cytochromes P-450,” Biochemical Pharmacology, Vol. 34, No. 18, 1985, pp. 3337- 3345. doi:10.1016/0006-2952(85)90355-7
W. Lilienblum, A. K. Walli and K. W. Bock, “Differential Induction of Rat Liver Microsomal UDP-Glu- curonosyl-Transferase Activites by Various Inducing Agents,” Biochemical Pharmacology, Vol. 31, No. 6, 1982, pp. 907-913. doi:10.1016/0006-2952(82)90319-7
J. Plazar, B. ?egura, T. T. Lah and M. Filipi?, “Protective Effects of Xanthohumol against the Genotoxicity of Benzo(a)pyrene (BaP), 2-Amino-3-Methylimidazo [4,5-f] Quinoline (IQ) and Tert-Butyl Hydroperoxide (t-BOOH) in HepG2 Human Hepatoma Cells,” Mutation Research/ Genetic Toxicology and Environmental Mutagenesis, Vol. 632, No. 1-2, 2007, pp. 1-8. doi:10.1016/j.mrgentox.2007.03.013
T. Sugimura, “Food and Cancer,” Toxicology, Vol. 181- 182, 2002, pp. 17-21. doi:10.1016/S0300-483X(02)00250-0
M. Murata, M. Kobayashi and S. Kawanishi, “Mechanism of Oxidative DNA Damage Induced by a Heterocyclic Amine, 2-Amino-3,8-Dimethylimidazo[4,5-F]Qui- noxaline,” Cancer Science, Vol. 90, No. 3, 1999, pp. 268- 275. doi:10.1111/j.1349-7006.1999.tb00743.x
J. H. Weisburger, “Comments on the History and Importance of Aromatic and Heterocyclic Amines in Public Health,” Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, Vol. 506-507, No. 9, 2002, pp. 9-20. doi:10.1016/S0027-5107(02)00147-1
F. T. Hatch, M. G. Knize and M. E. Colvin, “Extended Quantitative Structure-Activity Relationships for 80 Aromatic and Heterocyclic Amines: Structural, Electronic, and Hydropathic Factors Affecting Mutagenic Potency,” Environmental and Molecular Mutagenesis, Vol. 38, No. 4, 2001, pp. 268-291. doi:10.1002/em.10028
S. De Flora and L. R. Ferguson, “Overview of Mechanisms of Cancer Chemopreventive Agents,” Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, Vol. 591, No. 1-2, 2005, pp. 8-15. doi:10.1016/j.mrfmmm.2005.02.029
P. Dolara, C. Luceri, C. D. Filippo, A. P. Femia, L. Giovannelli, G. Caderni, C. Cecchini, S. Silvi, C. Orpianesi and A. Cresci, “Red Wine Polyphenols Influence Carcinogenesis, Intestinal Microflora, Oxidative Damage and Gene Expression Profiles of Colonic Mucosa in F344 Rats,” Mutation Research, Vol. 591, No. 1-2, 2005, pp. 237-246. doi:10.1016/j.mrfmmm.2005.04.022
J. D. Lambert, J. Hong, G. Yang, J. Liao and C. S. Yang, “Inhibition of Carcinogenesis by Polyphenols: Evidence from Laboratory Investigations,” The American Journal of Clinical Nutrition, Vol. 81, No. 1, 2005, pp. S284-S291.
K. Kanazawa, M. Uehara, H. Yanagitani and T. Hashimoto, “Bioavailable Flavonoids to Suppress the Formation of 8-OHdG in HepG2 Cells,” Archives of Bioche- mistry and Biophysics, Vol. 455, No. 2, 2006, pp. 197- 203. doi:10.1016/j.abb.2006.09.003
C. Rice-Evans, N. Miller and G. Paganga, “Structure- Antioxidant Activity Relationships of Flavonoids and Phenolic Acids,” Free Radical Biology & Medicine, Vol. 20, No. 7, 1996, pp. 933-956. doi:10.1016/0891-5849(95)02227-9
P. Makena and K. Chung, “Effects of Various Plant Polyphenols on Bladder Carcinogen Benzidine-Induced Mutagenicity,” Food and Chemical Toxicology, Vol. 45, No. 10, 2007, pp. 1899-1909. doi:10.1016/j.fct.2007.04.007
J. H. Weisburger and F. Chung, “Mechanisms of Chronic Disease Causation by Nutritional Factors and Tobacco Products and Their Prevention by Tea Polyphenols,” Food and Chemical Toxicology, Vol. 40, No. 8, 2002, pp. 1145-1154. doi:10.1016/S0278-6915(02)00044-3
F. T. Hatch, F. C. Lightstone and M. E. Colvin, “Quantitative Structure-Activity Relationship of Flavonoids for Inhibition of Heterocyclic Amine Mutagenicity,” Envi- ronmental and Molecular Mutagenesis, Vol. 35, No. 4, 2000, pp. 279-299. doi:10.1002/1098-2280(2000)35:4 3.0.CO;2-9
Z. Apostolides, D. A. Balentine, M. E. Harbowy, Y. Hara and J. H. Weisburger, “Inhibition of PhIP Mutagenicity by Catechins, and by Theaflavins and Gallate Esters,” Mutation Research/Genetic Toxicology and Environmen- tal Mutagenesis, Vol. 389, No. 2-3, 1997, pp. 167-172. doi:10.1016/S1383-5718(96)00143-X
P. Hodek, P. Trefil and M. Stiborová, “Flavonoids-Potent and Versatile Biologically Active Compounds Interacting with Cytochromes P450,” Chemico-Biological Interactions, Vol. 139, No. 1, 2002, pp. 1-21. doi:10.1016/S0009-2797(01)00285-X
J. Hümmerich, C. Zohm and W. Pfau, “Modulation of Cytochrome P450 1A1 by Food-Derived Heterocyclic Aromatic Amines,” Toxicology, Vol. 199, No. 2-3, 2004, pp. 231-240. doi:10.1016/j.tox.2004.02.028
R. J. Williams, J. P. E. Spencer and C. Rice-Evans, “Flavonoids: Antioxidants or Signalling Molecules?” Free Radical Biology and Medicine, Vol. 36, No. 7, 2004, pp. 838-849. doi:10.1016/j.freeradbiomed.2004.01.001
Y. J. Moon, X. Wang and M. E. Morris, “Dietary Flavonoids: Effects on Xenobiotic and Carcinogen Metabolism,” Toxicology in Vitro, Vol. 20, No. 2, 2006, pp. 187-210. doi:10.1016/j.tiv.2005.06.048