DOI QR코드

DOI QR Code

Genetic Toxicity Test of Emodin by Ames, Micronucleus, Comet Assays and Microarray Analysis Showing Differential Result

  • Go, Seo-Y. (College of Pharmacy, Ewha Womans University) ;
  • Kwon, Kyoung-J. (College of Pharmacy, Ewha Womans University) ;
  • Park, Sue-N. (Department of Toxicological Researches, National Institute of Toxicological Research, Korea Food and Drug Administration) ;
  • Sheen, Yhun-Y. (College of Pharmacy, Ewha Womans University)
  • 발행 : 2007.09.30

초록

Emodin (1,3,8-trihydroxy-6-methylanthraquinone) is a major constituent of rhubarb. Although it has been claimed to have a wild spectrum of therapeutic value, its side effects, especially in human kidney cells have not been well characterized. In this study, we have carried out in vitro genetic toxicity test of emodin and microarray analysis of differentially expressed genes in response to emodin. The result of Ames test showed mutations with emodin treatment in base substitution strain TA1535 both with and without exogenous metabolic activation. Likewise, emodin showed mutations in frame shift TA98 both with and without exogenous metabolic activation. The result of COMET assay in L5178Y cells with emodin treatment showed DNA damage both with and without exogenous metabolic activation. Emodin did not increase micronuclei in CHO cells both with and without exogenous metabolic activation. 150 Genes were selected as differentially expressed genes in response to emodin by microarray analysis and these genes would be candidate biomarkers of genetic toxic action of emodin.

키워드

참고문헌

  1. Affymetrix, Inc. 2000. '$GeneChip^{\circledR}$ Expression Analysis Technical Manual' http://www.affymetrix.com/support/technical /manual/expression_manual.affx
  2. Barnard D. L., Huffman J. H., Morris J. L., Wood S. G., Hughes B. G. and Sidwell R.W. (1992). Evaluation of the antiviral activity of anthraquinones, anthrones and anthraquinone derivatives against human cytomegalovirus. Antiviral Res. 17, 63-77
  3. Bosch R., Friederich U., Lutz W. K., Brocker E., Bachmann M. and Schlatter C. (1987). Investigations on DNA binding in rat liver and in Salmonella and on mutagenicity in the Ames test by emodin, a natural anthraquinone. Mutat. Res. 188, 161-168 https://doi.org/10.1016/0165-1218(87)90085-1
  4. Bruggeman I.M. and van der Hoeven J.C. (1984). Lack of activity of the bacterial mutagen emodin in HGPRT and SCE assay with V79 Chinese hamster cells. Mutat Res 138(2-3), 219-224 https://doi.org/10.1016/0165-1218(84)90047-8
  5. Brusick D. and Mengs U. (1997). Assessment of the genotoxic risk from laxative senna products. Environ. Mol. Mutagen. 29, 1-9 https://doi.org/10.1002/(SICI)1098-2280(1997)29:1<1::AID-EM1>3.0.CO;2-J
  6. Fenech M. (2000). The in vitro micronucleus technique. Mutat. Res 455, 81-95 https://doi.org/10.1016/S0027-5107(00)00065-8
  7. Gatehouse D., Haworth S., Cebula T., Gocke E., Kier L., Matsushima T., Melcion C., Nohmi T., Venitt S. and Zeiger E. (1994). Recommendations for the performance of bacterial mutation assays. Mutat. Res. 312, 217-233 https://doi.org/10.1016/0165-1161(94)90037-X
  8. Kasamatsu T., Ogura R., Ikeda N., Morita O., Saigo K., Watabe H., Saito Y. and Suzuki H. (2005). Genotoxicity studies on dietary diacylglycerol (DAG) oil. Food Chem. Toxicol. 43, 253-260 https://doi.org/10.1016/j.fct.2004.10.001
  9. Kirsch-Volders M., Sofuni T., Aardema M., Albertini S., Eastmond D., Fenech M., Ishidate M., Kirchner S., Lorge E., Morita T., Norppa H., Surralles J., Vanhauwaert A. and Wakata A. (2003). Report from the in vitro micronucleus assay working group. Mutat. Res. 540, 153-163 https://doi.org/10.1016/j.mrgentox.2003.07.005
  10. Kodama M., Kamioka Y., Nakayama T., Nagata C., Morooka N. and Ueno Y. (1987). Generation of free radical and hydrogen peroxide from 2-hydroxyemodin, a direct-acting mutagen, and DNA strand breaks by active oxygen. Toxicol. Lett. 37, 149-156 https://doi.org/10.1016/0378-4274(87)90151-2
  11. Krivobok S., Seigle-Murandi F., Steiman R., Marzin D. R. and Betina V. (1992). Mutagenicity of substituted anthraquinones in the Ames/Salmonella microsome system. Mutat. Res. 279, 1-8 https://doi.org/10.1016/0165-1218(92)90259-3
  12. Kumar A., Dhawan S. Aggarwal B. B. (1998). Emodin (3-methyl-1,6,8-trihydroxy anthraquinone) inhibits TNF-induced NF-kappaB activation, IkappaB degradation, and expression of cell surface adhesion proteins in human vascular endothelial cells. Oncogene 20, 17, 913-918
  13. Kuo Y. C., Meng H. C. and Tsai W. J. (2001). Regulation of cell proliferation, inflammatory cytokine production and calcium mobilization in primary human T lymphocytes by emodin from Polygonum hypoleucum Ohwi. Inflamm Res 50, 73-82 https://doi.org/10.1007/s000110050727
  14. Kuo Y. C., Sun C. M., Ou J. C. and Tsai W.J. (1997). A tumor cell growth inhibitor from Polygonum hypoleucum Ohwi. Life Sci. 61, 2335-2344 https://doi.org/10.1016/S0024-3205(97)00937-5
  15. Lee H. Z. (2001). Effects and mechanisms of emodin on cell death in human lung squamous cell carcinoma. Br. J. Pharmacol. 134, 11-20 https://doi.org/10.1038/sj.bjp.0704205
  16. Liu Z. H., Li L. S., Hu W. X. and Zhou H. (1996). Effect of emodin on c-myc proto-oncongen expression in cultured rat mesan-gial cells. Zhongguo Yao Li Xue Bao 17, 61-63
  17. Maclean W., Townsend P. Rhubarb (Da Huang): Rheum palmatum. http://www.acupunture.com/Herbology/Dahuang.htm. Accessed on June 7, 1999
  18. Masuda T., Haraikawa K., Morooka N. and Nakano S., (1985). Ueno Y. 2-Hydroxyemodin, an active metabolite of emodin in the hepatic microsomes of rats. Mutat. Res. 149, 327-332 https://doi.org/10.1016/0027-5107(85)90148-4
  19. Masuda T. and Ueno Y. (1984). Microsomal transformation of emodin into a direct mutagen. Mutat. Res. 125, 135-144 https://doi.org/10.1016/0027-5107(84)90065-4
  20. Mengs U., Krumbiegel G. and Volkner W. (1997). Lack of emodin genotoxicity in the mouse micronucleus assay. Mutat. Res. 393, 289-293 https://doi.org/10.1016/S1383-5718(97)00113-7
  21. Merck. 1998. The Merck Index Online. Dialog file 304. Emodin
  22. Mueller S. O., Lutz W. K., (1998). Stopper H. Factors affecting the genotoxic potency ranking of natural anthraquinones in mammalian cell culture systems. Mutat. Res. 414, 125-129 https://doi.org/10.1016/S1383-5718(98)00047-3
  23. Mueller S. O., Stopper H. (1999). Characterization of the genotoxicity of anthraquinones in mammalian cells. Biochim. Biophys. Acta. 1428, 406-414 https://doi.org/10.1016/S0304-4165(99)00064-1
  24. Muller S. O., Eckert I., Lutz W. K. and Stopper H. (1996). Genotoxicity of the laxative drug components emodin, aloe-emodin and danthron in mammalian cells: Topoisomerase II mediated? Mutat. Res. 371, 165-173 https://doi.org/10.1016/S0165-1218(96)90105-6
  25. Murakami H., Kobayashi J., Masuda T., Morooka N. and Ueno Y. (1987). omega-Hydroxyemodin, a major hepatic metabolite of emodin in various animals and its mutagenic activity. Mutat. Res. 180, 147-153 https://doi.org/10.1016/0027-5107(87)90209-0
  26. National Toxicology Program, 2001. NTP toxicology and carcinogenesis studies of emodin (CAS NO. 518-82-1) feed studies in F344/N rats and B6C3F1 mice. Natl Toxicol. Program Tech. Rep. Ser. 493, 1-278
  27. National Toxicology Program. 2003. Report and study status database: Emodin (CAS No. 518-82-1). National Toxicology Program Home-page, http://ntp-server.niehs.nih.gov. Accessed February 12, 2003
  28. Sato M., Maulik G., Bagchi D. and Das D.K. (2000). Myocardial protection by protykin, a novel extract of trans-resveratrol and emodin. Free Radic. Res. 32, 135-144 https://doi.org/10.1080/10715760000300141
  29. Singh N.T., McCoy M.T., Tice R.R. and Schneider E.L. (1988). A simple technique for quantification of low levels of DNA damage in individual cells. Exp. Cell. Res. 175, 184-191 https://doi.org/10.1016/0014-4827(88)90265-0
  30. von Wright A., Raatikainen O., Taipale H., Karenlampi S. and Mäki-Paakanen J. (1992). Directly acting geno-and cytotoxic agents from a wild mushroom Dermocybe sanguinea. Mutat. Res. 269, 27-33 https://doi.org/10.1016/0027-5107(92)90157-W
  31. Wehner F. C., Thiel P. G. and du Rand M. (1979). Mutagenicity of the mycotoxin emodin in the salmonella/microsome system. Appl. Environ. Microbiol. 37, 658-660
  32. Yeh S. F., Chou T. C. and Liu T. S. (1988). Effects of anthraquinones of Polygonum cuspidatum on HL-60 cells. Planta Med. 54, 413-414 https://doi.org/10.1055/s-2006-962484
  33. Zhang L. and Hung M. C. (1996). Sensitization of HER-2/neuoverexpressing non-small cell lung cancer cells to chemotherapeutic drugs by tyrosine kinase inhibitor emodin. Oncogene Feb 1; 12, 571-576
  34. Zhang L., Lau Y.K., Xi L., Hong R. L., Kim D. S., Chen C. F., Hortobagyi G. N., Chang C. and Hung M. C. (1998). Tyrosine kinase inhibitors, emodin and its derivative repress HER-2/neu-induced cellular transformation and metastasis-associated properties. Oncogene 4, 16, 2855-2863
  35. Zhang L., Lau Y. K., Xia W., Hortobagyi G. N. and Hung M. C. (1999). Tyrosine kinase inhibitor emodin suppresses growth of HER-2/neu-overexpressing breast cancer cells in athymic mice and sensitizes these cells to the inhibitory effect of paclitaxel. Clin. Cancer Res. 5, 343-353