Synergistic activation of estrogen receptor with combinations of environmental chemicals

Science

Volumn 272, Issue 5267, 1996, Pages 1489-1492

  Arnold, Steven F ^b   Klotz, Diane M ^b   Collins, Bridgette M ^b   Vonier, Peter M ^b   Guillette Jr , Louis J ^b   McLachlan, John A ^b  

^a TULANE UNIVERSITY SCHOOL OF MEDICINE   (United States)

^b NONE

Author keywords

[No Author keywords available]

Indexed keywords

CAMPHECLOR; DIELDRIN; ENDOSULFAN; ESTROGEN RECEPTOR; POLYCHLORINATED BIPHENYL;

ARTICLE; CONTROLLED STUDY; ESTROGEN ACTIVITY; NONHUMAN; POLLUTANT; PRIORITY JOURNAL; SEXUAL DEVELOPMENT; TRANSACTIVATION; TURTLE; YEAST;

EID: 0029972911     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.272.5267.1489     Document Type: Article

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21. 3H]17β-estradiol was measured by scintillation counting. The data shown are representative of at least three independent experiments (D. M. Klotz, J. A. McLachlan, S. F. Arnold, in preparation).
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32. Ishikawa cells grown in a 35-mm plate were transfected with 2 μg of pERE-luc (containing two copies of the vitellogenin estrogen response element linked to the luciferase gene), 1 μg of a control β-Gal plasmid, and 20 ng of hER in a mammalian expression vector with 10 μl of lipofectamine (GIBCO BRL) for 5 hours. The cells were treated with vehicle, estradiol, or 2′,4′,6′-trichloro-4-biphenylol or 2′,3′,4′,5′-tetraohloro-4-biphenylol (or both) in Dulbecco's modified Eagle's medium with 5% charcoal-stripped fetal bovine serum for 18 hours. All chemicals were dissolved in DMSO and added to the media so that the concentration of DMSO did not exceed 1%. The cells were collected by incubation with lysis buffer (Analytical Luminescence, Ann Arbor, MI) for 15 min at 25°C, the protein concentration determined with the Bio-Rad protein assay reagent, and the β-Gal activity measured as described (18). The amount of extract used in the luciferase assay was normalized for protein and β-Gal activity. The luciferase assay was performed in the Monolight 2010 as recommended by the manufacturer (Analytical Luminescence).
33. We thank Tulane University, the W. Alton Jones Foundation, and the Louisiana Breast Cancer Foundation for support; W. A. Toscano, N. Masahiko, and J. R. Ostberg for critical reading of the manuscript; and M. K. Robinson for help with the YES and S. Safe for providing the PCBs.