Steroid Hormone Receptor/Reporter Gene Transcription Assay for Food Additives and Contaminants

  • Jeong Sang-Hee (Toxicology Division, National Veterinary Research and Quarantine Service) ;
  • Cho Joon-Hyoung (Toxicology Division, National Veterinary Research and Quarantine Service) ;
  • Park Jong-Myung (Toxicology Division, National Veterinary Research and Quarantine Service)
  • 발행 : 2006.03.01

초록

Many of endocrine disrupting chemicals induce effects via interaction with hormone receptors and responsive elements in target cells. We investigated endocrine disrupting effects of some food additives and contaminants including BHA, BHT, ethoxyquin, propionic acid, sorbic acid, benzoic acid, CPM, aflatoxin B1, cadmium chloride, genistein, TCDD and PCBs in yeast transformants expressing human steroid hormone receptors along with steroid responsive elements. The response limit of genetically recombinant yeast to $17{\beta}$-estradiol, testosterone and progesterone was $1{\times}10^{-16},\;1{\times}10^{-12}\;and\;1{\times}10^{-13}M$, respectively. BHT induced weak transcriptional activity in estrogen sensitive yeast, while BHA and sorbic acid interacted weakly with androgen receptor/responsive element. CPM induced transcriptional activities in all types of yeasts sensitive to steroid hormones. Zearalenone and genistein induced high transcriptional activation in estrogen sensitive yeast with relative potencies almost $10^8$ folds lower than $17{\beta}$-estradiol. TCDD induced transcriptional activation weakly in estrogen- and progesterone- sensitive yeasts. This study elucidated that recombinant yeast is a sensitive and high-throughput system and can be used for the direct assessment on chemical interactions with steroid receptors and responsive elements. Also, the present study raises the requirement of evaluation on the endocrine disrupting effects of BHT, BHA, sorbic acid, CPM and TCDD for their transcription activity in yeast screening system though weak in intensity.

키워드

참고문헌

  1. Arnold, S.F., Klotz, D.M., Collins, B.M., Vonier, P.M., Guillette, L.J. Jr. and McLachlan, J.A. (1996): Synergistic activation of estrogen receptor with combinations of environmental chemicals. Science, 272, 1489-1491 https://doi.org/10.1126/science.272.5267.1489
  2. Ausubel, FM., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. and Struhl, K. (2000): Introduction of DNA into mammalian cells in Current protocols in molecular biology. Volume 2. John Wieley & Sons, Inc., USA, pp. 9.0.1-9.17.2
  3. Baker, V.A. (2001): Endocrine disrupters-testing strategies to assess human hazard. Toxicol. In Vitro, 15, 413-419 https://doi.org/10.1016/S0887-2333(01)00045-5
  4. Crisp, T.M., Clegg, E.D., Cooper, R.L., Wood, W.P., Anderson, D.G., Baetcke, K.P., Hoffmann, J.L., Morrow, M.S., Rodier, D.J., Schaeffer, J.E., Touart, L.W., Zeeman, M.G. and Patel, Y.M. (1998): Environmental endocrine disruption: an effects assessment and analysis. Environ. Health Perspect., 106, Suppl. 11-56 https://doi.org/10.2307/3433911
  5. Danzo, B.J. (1997): Environmental xenobiotics may disrupt normal endocrine function by interfering with the binding of physiological ligands to steroid receptors and binding proteins. Environ. Health Perspect., 105, 294-301 https://doi.org/10.2307/3433266
  6. Daston, G.P., Cook, J.C. and Kavlock, R.J. (2003): Uncertainties for endocrine disrupters: our view on progress. Toxicol. Sci., 74, 245-252 https://doi.org/10.1093/toxsci/kfg105
  7. Desaulniers, D., Poon, R., Phan, W., Leingartner, K., Foster, W.G. and Chu, I. (1997): Reproductive and thyroid hormone levels in rats following 90-day dietary exposure to PCB 28 (2,4,4'-trichlorobiphenyl) or PCB 77 (3,3',4,4'-tetrachlorobiphenyl). Toxicol. Ind. Health, 13, 627-638 https://doi.org/10.1177/074823379701300504
  8. Gaido, K.W., Leonard, L.S., Lovell, S., Gould, J.C., Babai, D., Portier, C.J. and McDonnell, D.P. (1997): Evaluation of chemicals with endocrine modulating activity in a yeastbased steroid hormone receptor gene transcription assay. Toxicol. Appl. Pharmacol., 143, 205-212 https://doi.org/10.1006/taap.1996.8069
  9. Gray, L.E. Jr., Kelce, W.R., Wiese, T., Tyl, R., Gaido, K., Cook, J., Klinefelter, G., Desaulniers, D., Wilson, E., Zacharewski, T., Waller, C., Foster, P., Laskey, J., Reel, J., Giesy, J., Laws, S., McLaclan, J., Breslin, W., Cooper, R., Di Giulio, R., Johnson, R., Purdy, R., Mihaich, E., Safe, S., Sonnenschein, C., Welshons, W., Miller, R., McMaster, S. and Colborn, T. (1997): Endocrine screening methods workshop report: detection of estrogenic and adrogenic hormonal and antihormonal activity for chemicals that act via receptor or steroidogenic enzyme mechanisms. Reprod. Toxicol., 11, 719-750 https://doi.org/10.1016/S0890-6238(97)00025-7
  10. Gutendorf, B. and Westendorf, J. (2001): Comparison of an array of in vitro assays for the assessment of the estrogenic potential of natural and synthetic estrogens, phytoestrogens and xenoestrogens. Toxicology, 166, 79-89 https://doi.org/10.1016/S0300-483X(01)00437-1
  11. IEH (1999): IEH assessment on the ecological significance on endocrine disruption. Page Bros, Norwich, UK, pp. 31-38
  12. Jeong, S.H., Kim, B.Y., Kang, H.G., Ku, H.O. and Cho, J.H. (2005a): Effects of butylated hydroxyanisole on the development and functions of reproductive system in rats. Toxicology, 208, 49-62 https://doi.org/10.1016/j.tox.2004.11.014
  13. Jeong, S.H., Kim, B.Y., Kang, H.G., Ku, H.O. and Cho, J.H. (2005b): Effect of chlorpyrifos-methyl on steroid and thyroid hormones in rat F0- and F1-generations. Toxicology, 216, in press
  14. Johnson, F.M. (2002): How many food additives are rodent carcinogens? Environ. Mol. Mutagen., 39, 69-80 https://doi.org/10.1002/em.10037
  15. Kang, H.G., Jeong, S.H., Cho, J.H., Kim. D.G., Park, J.M. and Cho, M.H. (2004): Chlorpyrifos-methyl shows anti-androgenic activity without estrogenic activity in rats. Toxicology, 199, 219-230 https://doi.org/10.1016/j.tox.2004.02.025
  16. Kang, H.G., Jeong, S.H., Cho, J.H., Kim, D.G., Park, J.M. and Cho, M.H. (2005): Evaluation of estrogenic and androgenic activity of butylated hydroxyanisole in immature female and castrated rats. Toxicology, 213, 147-156 https://doi.org/10.1016/j.tox.2005.05.027
  17. Klotz, D.M., Beckman, B.S., Hill, S.M., McLachlan, J.A., Walters, M.R. and Arnold, S.F. (1996): Identification of environmental chemicals with estrogenic activity using a combination of in vitro assays. Envirion. Health Perspect., 104, 1084-1089 https://doi.org/10.2307/3433121
  18. Kohn, M.C., Sewall, C.H., Lucier, G.W. and Portier, C.J. (1996): A mechanistic model of effects of dioxin on thyroid hormones in the rat. Toxicol. Appl. Pharmacol., 136, 29-48 https://doi.org/10.1006/taap.1996.0004
  19. Korner, W., Hanf, V., Schuller, W., Bartsch, H., Zwirner, M. and Hagenmaier H. (1998): Validation and application of a rapid in vitro assay for assessing the estrogenic potency of halogenated phenolic chemicals. Chemosphere, 37, 2395-2407 https://doi.org/10.1016/S0045-6535(98)00297-5
  20. McLachlan, J.A. (1993): Functional toxicology: A new approach to detect biologically active xenobiotics. Environ. Health Perspect., 101, 386-387 https://doi.org/10.2307/3431891
  21. Nilsson, R. (2000): Endocrine modulators in the food chain and environment. Toxicol. Pathol., 28, 420-431 https://doi.org/10.1177/019262330002800311
  22. O'Connor, J.C., Cook, J.C., Slone, T.W., Makovec, G.T., Frame, S.R. and Davis, L.G. (1998): An ongoing validation of a Tier I screening battery for detecting endocrine-active compounds (EACs). Toxicol. Sci., 46, 45-60 https://doi.org/10.1006/toxs.1998.2550
  23. O'Connor, J.C., Davis, L.G., Frame, S.R. and Cook, J.C. (2002): Evaluation of a Tier I screening battery for detecting endocrine-active compounds (EACs) using the positive controls testosterone, coumestrol, progesterone, and RU486. Toxicol. Sci., 54, 338-354 https://doi.org/10.1093/toxsci/54.2.338
  24. Osius, N., Karmaus, W., Kruse, H. and Witten, J. (1999): Exposure to polychlorinated biphenyls and levels of thyroid hormones in children. Environ. Health Perspect., 107, 843-849 https://doi.org/10.2307/3454584
  25. Pham, T.A., Hwung, Y.P., Santiso-Mere, D., McDonnell, D.P. and O'Malley, B.W. (1992): Ligand-dependent and-independent function of the transactivation regions of the human estrogen receptor in yeast. Mol. Endocrinol., 6, 1043-1050 https://doi.org/10.1210/me.6.7.1043
  26. Roy, D., Palangat, M., Chen, C.W., Thomas, R.D., Colerangle, J., Atkinson, A. and Yan, Z.J. (1997): Biochemical and molecular changes at the cellular level in response to exposure to environmental estrogen-like chemicals. J. Toxicol. Environ. Health, 50, 1-29 https://doi.org/10.1080/009841097160573
  27. Sonnenschein, C. and Soto, A.M. (1998): An updated review of environmental estrogen and androgen mimics and antagonists. J. Steroid Biochem. Mol. Biol., 65, 143-150 https://doi.org/10.1016/S0960-0760(98)00027-2
  28. Takeyoshi, M., Yamasaki, K., Sawaki, M., Nakai, M., Noda, S. and Takatsuki, M. (2002): The efficacy of endocrine disruptor screening tests in detecting anti-estrogenic effects downstream of receptor-ligand interactions. Toxicol. Lett., 126, 91-98 https://doi.org/10.1016/S0378-4274(01)00446-5
  29. Vegeto, E., Allan, G.F., Schrader, W.T., Tsai, M.J., McDonnell, D.P. and O'Malley, B.W. (1992): The mechanism of RU486 antagonism is dependent on the conformation of the carboxy- terminal tail of the human progesterone receptor. Cell, 69, 703-713 https://doi.org/10.1016/0092-8674(92)90234-4
  30. Wang, T.T., Sathyamoorthy, N. and Phang, J.M. (1996): Molecular effects of genistein on estrogen receptor mediated pathways. Carcinogenesis, 17, 271-275 https://doi.org/10.1093/carcin/17.2.271