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Transfected HepG2 Cells for Evaluation of Catechin Effects on Alcohol-Induced CYP2E1 Cytotoxicity  

LEE YOO-HYUN (School of Life Sciences & Biotechnology, Korea University)
HO JIN-NYOUNG (School of Life Sciences & Biotechnology, Korea University)
DONG MI-SOOK (School of Life Sciences & Biotechnology, Korea University)
PARK CHANG-HWAN (College of Medicine, Hanyang University)
KIM HYE-KYUNG (Department of Food and Biotechnology, Hanseo University)
HONG BUMSHIK (School of Life Sciences & Biotechnology, Korea University)
SHIN DONG-HOON (School of Life Sciences & Biotechnology, Korea University)
CHO HONG-YON (School of Life Sciences & Biotechnology, Korea University)
Publication Information
Journal of Microbiology and Biotechnology / v.15, no.6, 2005 , pp. 1310-1316 More about this Journal
Abstract
To evaluate the toxicological properties of human cytochrome P450 2E1 (CYP2E1) induced by ethanol and possible protective effects of various green tea catechins on alcohol-induced toxicity, transfected HepG2 cells that stably and constitutively express human CYP2E1 were established using the recombinant retroviral expression vector. Exposure of the CYP2E1-expressing HepG2 cells to high concentration of ethanol (200 mM) for 5 days resulted in a more than $50\%$ increase of cytotoxicity, assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction, lactate dehydrogenase (LDH) release, and reactive oxygen species (ROS) production, and loss of normal morphology, in comparison with HepG2 cells containing control vector. Treatment of the cells with various catechins increased cell viability by more than 2-fold. (-)-Epicatechin gallate and(-)-catechin gallate at the lowest concentration ($5\;{\mu}M$) attenuated cell death induced CYP2E1 by $60-65\%$. Therefore, the results showed that the catechins, including epimerized catechins, have strong protective effects against alcohol-induced CYP2E1 toxicity, and it is correlated with antioxidant effect.
Keywords
HepG2 cells; cytochrome P450 2E1; ethanol; catechins;
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1 Aguilar, C. N., M. Cruz, R. Rodriguez, G. Gutierrenz- Sanchez, A. Ramirez-Coronel, and C. Augur. 2004. Catechin degradation by several fungal strains isolated from Mexican desert. J. Microbiol. Biotechnol. 14: 426-429
2 Chen, Q. and A. I. Cederbaum. 1998. Cytotoxicity and apoptosis produced by cytochrome P450 2E1 in HepG2 cells. Mol. Pharmacol. 53: 638-648   DOI
3 Donohue, T. M., D. L. Clemens, Jr., A. Galli, D. Crabb, N. Nieto, J. Kato, and S. S. Barve. 2001. Use of cultured cells in assessing ethanol toxicity and ethanol-related metabolism. Alcohol Clin. Exp. Res. 25: 87S-93S   DOI   ScienceOn
4 Eysseric, H., B. Gonthier, A. Soubeyran, M. J. Richard, D. Daveloose, and L. Barret. 2000. Effects of chronic ethanol exposure on acetaldehyde and free radical production by astrocytes in culture. Alcohol 21: 117-125   DOI   ScienceOn
5 Ikeda, I., M. Kobayashi, T. Hamada, K. Tsuda, H. Goto, K. Imaizumi, A. Nozawa, A. Sugimoto, and T. Kakuda. 2003. Heat-epimerized tea catechins rich in gallocatechin gallate and catechins gallate are more effective to inhibit cholesterol absorption than tea catechins rich in epigallocatechin gallate and epicatechin gallate. J. Agric. Food Chem. 51: 7303- 7307   DOI   ScienceOn
6 Kurose, I., H. Higuchi, S. Miura, H. Saito, N. Watanabe, R. Hokari, M. Hirokawa, M. Takaishi, S. Zeki, T. Nakamura, H. Ebinuma, S. Kato, and H. Ishii. 1997. Oxidative stressmediated apoptosis of hepatocytes exposed to acute ethanol intoxication. Hepatology 25: 368-378   DOI   ScienceOn
7 Muto, S., K. Fujita, Y. Yamazak, and T. Kamatak. 2001. Inhibition by green tea catechins of metabolic activation of procarcinogens by human cytochrome P450. Mutat. Res. 479: 197-206   DOI
8 Nouso, K., S. S. Thorgeirsson, and N. Battula. 1992. Stable expression of human cytochrome P450IIE1 in mammalian cells: Metabolic activation of nitrosodimethylamine and formation of adducts with cellular DNA. Cancer Res. 52: 1796-1800
9 Xu, Y., M. A. Leo, and C. S. Lieber. 2003. Lycopene attenuates arachidonic acid toxicity in HepG2 cells overexpressing CYP2E1. Biochem. Biophys. Res. Commun. 303: 745-750   DOI   ScienceOn
10 Wu, D. and A. I. Cederbaum. 1996. Ethanol cytotoxicity to a transfected HepG2 cell line expressing human cytochrome P4502E1. J. Biol. Chem. 271: 23914-23919   DOI   ScienceOn
11 Cameron, R. G., M. G. Neuman, N. H. Shear, G. Katz, S. Bellentani, and C. Tiribelli. 1998. Modulation of liver-specific cellular response to ethanol in vitro in HepG2 cells. Toxicol. In Vitro 12: 111-122   DOI   ScienceOn
12 Lee, H. J. and M. B. Gu. 2003. Effect of benzo[a]pyrene on genes related to the cell cycle and cytochrome P450 of Saccharomyces cerevisiae. J. Microbiol. Biotechnol. 13: 624-627
13 Castilla, R., R. Gonzalez, D. Fouad, E. Fraga, and J. Muntane. 2004. Dual effect of ethanol on cell death in primary culture of human and rat hepatocytes. Alcohol Alcohol. 39: 290- 296   DOI   ScienceOn
14 Kagaya, N., Y. Tagawa, H. Nagashima, R. Saijo, M. Kawase, and K. Yaki. 2002. Suppression of cytotoxin-induced cell death in isolated hepatocytes by tea catechins. Eur. J. Pharmacol. 450: 231-236   DOI   ScienceOn
15 Ostrowska, J., W. Luczaj, I. Kasacka, A. Rozanski, and E. Skrzydlewska. 2004. Green tea protects against ethanolinduced lipid peroxidation in rat organs. Alcohol 32: 25-32   DOI   ScienceOn
16 Jamenez-Lopez, J. M. and A. I. Cederbaum. 2003. Green tea polyphenol epigallocatechin-3-gallate protects HepG2 cells against CYP2E1-dependent toxicity. Free Radic. Biol. Med. 36: 359-370   DOI   ScienceOn
17 Kim, S., J. S. Kang, D. D. Jang, K. Lee, S. A. Kim, B. S. Han, and Y. I. Park. 2004. Differential gene expression in estradiol-3-benzoate-treated liver and chemically-induced hepatocellular carcinoma. J. Microbiol. Biotechnol. 14: 1286-1294
18 Lee, S.-K., C.-Y. Choi, J.-S. Ahn, J.-Y. Cho, C.-S. Park, and Y. J. Yoon. 2004. Identification of a cytochrome P450 hydroxylation gene involved in rifamycin biosynthesis by Amycolatopsis mediterranei S699. J. Microbiol. Biotechnol. 14: 356-362
19 Mapoles, J., F. Berthou, A. Alexander, F. Simon, and J. F. Menez. 1993. Mammalian PC-12 cell genetically engineered for human cytochrome P450 2E1 expression. Eur. J. Biochem. 214: 735-745   DOI   ScienceOn
20 Sun, A. Y., M. Ingelman-Sundberg, E. Neve, H. Matsumoto, Y. Nishitani, Y. Minowa, Y. Fukui, S. M. Bailey, V. B. Patel, C. C. Cunningham, T. Zima, L. Fialova, L. Mikulikova, P. Popov, I. Malbohan, M. Janebova, K. Nespor, and G. Y. Sun. 2001. Ethanol and oxidative stress. Alcohol Clin. Exp. Res. 25: 237S-243S   DOI   ScienceOn
21 Caro, A. A. and A. I. Cederbaum. 2004. Oxidative stress, toxicology, and pharmacology of CYP2E1. Annu. Rev. Pharmacol. Toxicol. 44: 27-42   DOI   ScienceOn
22 Ory, D. S., B. A. Neugeboren, and R. C. Mulligan. 1996. A stable human-derived packaging cell line for production of high titer retrovirus/vesicular stomatitis virus G pseudotypes. Proc. Natl. Acad. Sci. USA 93: 11400-11406
23 Zhuge, J., Y. Luo, and Y. N. Yu. 2003. Heterologous expression of human cytochrome P450 2E1 in HepG2 cell line. World J. Gastroentrol. 9: 2732-2736   DOI
24 Li, Y., D. W. Walker, and M. A. King. 2001. Peroxide mediates ethanol-induced cytotoxicity in PC12 cells. Free Radic. Biol. Med. 30: 389-392   DOI   ScienceOn
25 Waxman, D. J., D. P. Lapenson, T. Aoyama, H. V. Gelboin, F. J. Gonzalez, and K. Korzekwa. 1991. Steroid hormone hydroxylase specificities of eleven cDNA-expressed human cytochrome P450s. Arch. Biochem. Biophys. 290: 160-166   DOI   ScienceOn
26 Xu, Y., M. A. Leo, and C. S. Lieber. 2003. Lycopene attenuates alcoholic apoptosis in HepG2 cells expressing CYP2E1. Biochem. Biophys. Res. Commun. 308: 614-618   DOI   ScienceOn
27 Bailey, S. M. and C. C. Cunningham. 1999. Acute and chronic ethanol increases reactive oxygen species generation and decreases viability in fresh, isolated rat hepatocytes. Hepatology 28: 1318-1326
28 Lamarche, F., B. Gonthier, N. Signorini, H. Eysseric, and L. Baret. 2003. Acute exposure of cultured neurons to ethanol results in reversible DNA single-stranded breaks, whereas chronic exposure causes loss of cell viability. Alcohol Alcohol. 38: 550-558   DOI   ScienceOn
29 Balentine, D. A. and I. Paetau-Robinson. 1998. Tea as a source of dietary antioxidants with a potential role in prevention of chronic diseases, pp. 265-287. In G. Mazza and B. D. Oomah (eds.), Herbs, Botanicals, and Teas. CRC Press LLC, Florida
30 Chen, L., X. Yang, H. Jiao, and B. Zhao. 2002. Tea catechins protect against lead-induced cytotoxicity, lipid peroxidation, and membrane fluidity in HepG2 cells. Toxicol. Sci. 69: 249-256
31 Cederbaum, A. I. 2003. Iron and CYP2E1-dependent oxidative stress and toxicity. Alcohol 30: 115-120   DOI   PUBMED   ScienceOn
32 Lieber, C. S. 2000. Alcohol: Its metabolism and interaction with nutrients. Annu. Rev. Nutr. 20: 395-430   DOI   PUBMED   ScienceOn
33 Wu, D. and A. I. Cederbaum. 1999. Ethanol-induced apoptosis to stable HepG2 cell lines expressing human cytochrome P-4502E1. Alcohol Clin. Exp. Res. 23: 67-76   DOI   ScienceOn
34 Kim, J.-H., H.-J. Kim, J.-H. Son, H.-N. Chun, J.-O. Yang, S.-J. Choi, N.-S. Paek, G.-H. Choi, and S.-K. Kim. 2003. Effect of Lactobacillus fermentum MG 590 on alcohol metabolism and liver function in rats. J. Microbiol. Biotechnol. 13: 919-925
35 Dai, Y., J. Rashba-Step, and A. I. Cederbaum. 1993. Stable expression of human P4502E1 in HepG2 cells: Characterization of catalytic activities and production of reactive oxygen intermediates. Biochemistry 32: 6928-6937   DOI   ScienceOn
36 Neuman, M. G., G. Koren, and C. Tribelli. 1993. In vitro assessment of the ethanol-induced hepatotoxicity in HepG2 cell line. Biochem. Biophys. Res. Commun. 197: 932-941   DOI   ScienceOn
37 Nakagawa, T., M. Sawada, F. J. Gonzalez, T. Yokoi, and T. Kamaraki. 1996. Stable expression of human CYP2E1 in Chinese hamster cells: High sensitivity to N,N-dimethylnitrosamine in cytotoxicity testing. Mutat. Res. 360: 181-186   DOI   ScienceOn