Detection of estrogenic activity in sediment-associated compounds using in vitro reporter gene assays

Detection of estrogenic activity in sediment-associated compounds using in vitro reporter gene... Sediments may be the ultimate sink for persistent (xeno-)estrogenic compounds released into the aquatic environment. Sediment-associated estrogenic potency was measured with an estrogen receptor-mediated luciferase reporter gene (ER-CALUX) assay and compared with a recombinant yeast screen. The ER-CALUX assay was more sensitive to 17β-estradiol (E2) than the recombinant yeast screen, with an EC50 of 6 pM E2 compared to 100 pM in the yeast screen. Yeast cells were unable to distinguish the anti-estrogens ICI 182 780 and (4-hydroxy)tamoxifen, which were agonistic in the yeast. Acetone-soluble fractions of hexane/acetone extracts of sediments showed higher estrogenic potency than hexane-soluble extracts in the ER-CALUX assay. Sediments obtained from industrialized areas such as the Port of Rotterdam showed the highest estrogenic potency of the 12 marine sediments tested (up to 40 pmol estradiol equivalents per gram sediment). The estrogenic activity of individual chemicals that can be found in sediments including: alkylphenol ethoxylates and carboxylates; phthalates; and pesticides, was tested. Increasing sidechain length of various nonylphenol ethoxylates resulted in decreased estrogenic activity. Of the phthalates tested, butylbenzylphthalate was the most estrogenic, though with a potency approximately 100 000 times less than E2. The organochlorine herbicides atrazine and simazine failed to induce reporter gene activity. As metabolic activation may be required to induce estrogenic activity, a metabolic transformation step was added to the ER-CALUX assay using incubation of compounds with liver microsomes obtained from PCB-treated rats. Results indicate that metabolites of E2, NP and bisphenol A were less active than the parent compounds, while metabolites of methoxychlor were more estrogenic following microsomal incubations. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Science of the Total Environment Elsevier

Detection of estrogenic activity in sediment-associated compounds using in vitro reporter gene assays

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Publisher
Elsevier
Copyright
Copyright © 2002 Elsevier Science B.V.
ISSN
0048-9697
eISSN
1879-1026
D.O.I.
10.1016/S0048-9697(01)01146-9
Publisher site
See Article on Publisher Site

Abstract

Sediments may be the ultimate sink for persistent (xeno-)estrogenic compounds released into the aquatic environment. Sediment-associated estrogenic potency was measured with an estrogen receptor-mediated luciferase reporter gene (ER-CALUX) assay and compared with a recombinant yeast screen. The ER-CALUX assay was more sensitive to 17β-estradiol (E2) than the recombinant yeast screen, with an EC50 of 6 pM E2 compared to 100 pM in the yeast screen. Yeast cells were unable to distinguish the anti-estrogens ICI 182 780 and (4-hydroxy)tamoxifen, which were agonistic in the yeast. Acetone-soluble fractions of hexane/acetone extracts of sediments showed higher estrogenic potency than hexane-soluble extracts in the ER-CALUX assay. Sediments obtained from industrialized areas such as the Port of Rotterdam showed the highest estrogenic potency of the 12 marine sediments tested (up to 40 pmol estradiol equivalents per gram sediment). The estrogenic activity of individual chemicals that can be found in sediments including: alkylphenol ethoxylates and carboxylates; phthalates; and pesticides, was tested. Increasing sidechain length of various nonylphenol ethoxylates resulted in decreased estrogenic activity. Of the phthalates tested, butylbenzylphthalate was the most estrogenic, though with a potency approximately 100 000 times less than E2. The organochlorine herbicides atrazine and simazine failed to induce reporter gene activity. As metabolic activation may be required to induce estrogenic activity, a metabolic transformation step was added to the ER-CALUX assay using incubation of compounds with liver microsomes obtained from PCB-treated rats. Results indicate that metabolites of E2, NP and bisphenol A were less active than the parent compounds, while metabolites of methoxychlor were more estrogenic following microsomal incubations.

Journal

Science of the Total EnvironmentElsevier

Published: Jul 3, 2002

References

  • Issues arising when interpreting results from an in vitro assay for estrogenic activity
    Beresford, N.; Routledge, E.J.; Harris, C.A.; Sumpter, J.P.
  • Mutations in the yeast PDR3, PDR4, PDR7 and PDR9 pleiotropic (multiple) drug resistance loci affect the transcript level of and ATP binding cassette transporter encoding gene, PDR5
    Dexter, D.; Moye-Rowley, W.S.; Wu, A.L.; Golin, J.
  • Evaluation of chemicals with endocrine modulating activity in a yeast-based steroid hormone receptor gene transcription assay
    Gaido, K.W.; Leonard, L.S.; Lovell, S.; Gould, J.C.; Babai, D.; Portier, C.J.; McDonnell, D.P.
  • Detection of estrogenic potency in wastewater and surface water with three in vitro bioassays
    Murk, A.J.; Legler, J.; Van Lipzig, M.M.H.; Meerman, A.; Belfroid, A.C.; Spenkelink, A.; Rijs, G.B.J.; Vethaak, A.D.
  • Estrogenic activity of surfactants and some of their degradation products using a recombinant yeast screen
    Routledge, E.J.; Sumpter, J.P.
  • Activation of the human estrogen receptor by estrogenic and antiestrogenic compounds in Saccharomyces cerevisiae : a positive selection system
    Shiau, S.P.; Glasebrook, A.; Hardikar, S.D.; Yang, N.N.; Hershberger, C.L.
  • Functional role of estrogen metabolism in target cells: review and perspectives
    Zhu, B.T.; Conney, A.H.

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