Effect of Endocrine Disrupting Chemicals on Bombina orientalis Aromatase Activity Expressed in Cultured Mammalian Cells

  • Received : 2011.02.05
  • Accepted : 2011.02.15
  • Published : 2011.02.28

Abstract

In this study we present a mammalian cell culture model that allows to study the effect of endocrine disruptors (EDCs) on aromatase activity of aquatic amphibian, Bombina orientalis. Bombina orientalis aromatase gene was subcloned into a mammalian expression vector and subsequently transfected to mammalian cells. Although the protein expression level of Bombina orientalis aromatase was low, it had a significant aromatase activity. When EDCs were added to aromatase transfected cells, aromatase activity was significantly decreased. We report here that this system may be used to monitor the effect of EDCs on aromatase activity of aquatic organisms.

Keywords

References

  1. Ashby J, J Odum, D Paton, PA Lefevre, N Beresford and JP Sumpter. 2000. Re-evaluation of the first synthetic estrogen, 1-keto-1,2,3, 4-tetrahydrophenanthrene, and bisphenol A, using both the ovariectomised rat model used in 1933 and additional assays. Toxicol. Lett. 115:231-238. https://doi.org/10.1016/S0378-4274(00)00198-3
  2. Ashby J and H Tinwell. 1998. Uterotrophic activity of bisphenol A in the immature rat. Environ. Health Perspec. 106:719- 720. https://doi.org/10.1289/ehp.98106719
  3. Bonefeld-Jorgensen EC. 2004. The human health effect programme in Greenland, a review. Sci. Total Environ. 331: 215-231. https://doi.org/10.1016/j.scitotenv.2004.03.030
  4. Chang CY and E Witschi. 1956. Genic control and hormonal reversal of sex differentiation in Xenopus. Proc. Soc. Exp. Biol. Me. 93:140-144. https://doi.org/10.3181/00379727-93-22688
  5. Chang X, T Kobayashi, B Senthilkumaran, H Kobayashi-Kajura, CC Sudhakumari and Y Nagahama. 2005. Two types of aromatase with different encoding genes, tissue distribution and developmental expression in Nile tilapia (Oreochromis niloticus). Gen. Comp. Endocrinol. 141:101-115. https://doi.org/10.1016/j.ygcen.2004.11.020
  6. Cheshenko K, F Pakdel, H Segner, O Kah and RI Eggen. 2008. Interference of endocrine disrupting chemicals with aromatase CYP19 expression or activity, and consequences for reproduction of teleost fish. Gen. Comp. Endocrinol. 155: 31-62. https://doi.org/10.1016/j.ygcen.2007.03.005
  7. Gaido KW, LS Leonard, S Lovell, JC Gould, D Babai, CJ Portier and DP McDonnell. 1997. Evaluation of chemicals with endocrine modulating activity in a yeast-based steroid hormone receptor gene transcription assay. Toxicol. Appl. Pharmacol. 143:205-212. https://doi.org/10.1006/taap.1996.8069
  8. Graham FL and AJ van der Eb. 1973. Transformation of rat cells by DNA of human adenovirus 5. Virology 54:536-539. https://doi.org/10.1016/0042-6822(73)90163-3
  9. Kato T, K Matsui, M Takase, M Kobayashi and M Nakamura. 2004. Expression of P450 aromatase protein in developing and in sex-reversed gonads of the XX/XY type of the frog Rana rugosa. Gen. Comp. Endocrinol. 137:227-236. https://doi.org/10.1016/j.ygcen.2004.03.013
  10. Kitamura S, T Suzuki, S Sanoh, R Kohta, N Jinno, K Sugihara, S Yoshihara, N Fujimoto, H Watanabe and S Ohta. 2005. Comparative study of the endocrine-disrupting activity of bisphenol A and 19 related compounds. Toxicol. Sci. 84: 249-259. https://doi.org/10.1093/toxsci/kfi074
  11. Krishnan AV, P Stathis, SF Permuth, L Tokes and D Feldman. 1993. Bisphenol-A: an estrogenic substance is released from polycarbonate flasks during autoclaving. Endocrinology 132:2279-2286. https://doi.org/10.1210/en.132.6.2279
  12. Lee KM, W Yang, JS Rhee, DS Hwang, CJ Park, MC Gye, JS Lee and I Shin. 2010. Effects of endocrine disruptors on Bombina orientalis P450 aromatase activity. Zoolog. Sci. 27:338-343. https://doi.org/10.2108/zsj.27.338
  13. Lephart ED and ER Simpson. 1991. Assay of aromatase activity. Methods Enzymol. 206:477-483.
  14. Matthews JB, K Twomey and TR Zacharewski. 2001. In vitro and in vivo interactions of bisphenol A and its metabolite, bisphenol A glucuronide, with estrogen receptors alpha and beta. Chem. Res. Toxicol. 14:149-157. https://doi.org/10.1021/tx0001833
  15. Murthy JN, M Nagaraju, GM Sastry, AR Rao and GN Sastry. 2005. Active site acidic residues and structural analysis of modelled human aromatase: a potential drug target for breast cancer. J. Comput. Aided. Mol. Des. 19:857-870.
  16. Nagao T, K Wada, H Marumo, S Yoshimura and H Ono. 2001. Reproductive effects of nonylphenol in rats after gavage administration: a two-generation study. Reprod. Toxicol. 15:293-315. https://doi.org/10.1016/S0890-6238(01)00123-X
  17. Noaksson E, U Tjarnlund, AT Bosveld and L Balk. 2001. Evidence for endocrine disruption in perch (Perca fluviatilis) and roach (Rutilus rutilus) in a remote Swedish lake in the vicinity of a public refuse dump. Toxicol. Appl. Pharmacol. 174:160-176. https://doi.org/10.1006/taap.2001.9194
  18. Rotchell JM and GK Ostrander. 2003. Molecular markers of endocrine disruption in aquatic organisms. J. Toxicol. Environ. Health B Crit. Rev. 6:453-496. https://doi.org/10.1080/10937400306476
  19. Scholz S and HO Gutzeit. 2000. 17-alpha-ethinylestradiol affects reproduction, sexual differentiation and aromatase gene expression of the medaka (Oryzias latipes). Aquat. Toxicol. 50:363-373. https://doi.org/10.1016/S0166-445X(00)00090-4
  20. Soto AM, H Justicia, JW Wray and C Sonnenschein. 1991. p- Nonyl-phenol: an estrogenic xenobiotic released from "modified" polystyrene. Environ. Health Perspect. 92:167- 173. https://doi.org/10.1289/ehp.9192167
  21. Tinwell H, R Joiner, I Pate, A Soames, J Foster and J Ashby. 2000. Uterotrophic activity of bisphenol A in the immature mouse. Regul. Toxicol. Pharmacol. 32:118-126. https://doi.org/10.1006/rtph.2000.1412
  22. Weber LP, Y Kiparissis, GS Hwang, AJ Niimi, DM Janz and CD Metcalfe. 2002. Increased cellular apoptosis after chronic aqueous exposure to nonylphenol and quercetin in adult medaka (Oryzias latipes). Comp. Biochem. Physiol. C Toxicol. Pharmacol. 131:51-59. https://doi.org/10.1016/S1532-0456(01)00276-9
  23. Yung LY, ST Tsim and YH Wong. 1995. Stimulation of cAMP accumulation by the cloned Xenopus melatonin receptor through Gi and Gz proteins. FEBS Lett. 372:99-102. https://doi.org/10.1016/0014-5793(95)00963-A