Incorporation of S-9 activation into an ER-α transactivation assay

Grantley D. Charles; Michael J. Bartels; C. Gennings; Timothy R. Zacharewski; Nancy L. Freshour; B. Bhaskar Gollapudi; Edward W. Carney

doi:10.1016/S0890-6238(00)00070-8

Incorporation of S-9 activation into an ER-α transactivation assay

Grantley D. Charles, Michael J. Bartels, C. Gennings, Timothy R. Zacharewski, Nancy L. Freshour, B. Bhaskar Gollapudi, Edward W. Carney

Research output: Contribution to journal › Article › peer-review

21 Scopus citations

Abstract

We evaluated the feasibility of incorporating an exogenous metabolic activating system into an estrogen receptor-α transactivation assay. 17β- estradiol (E2), and the proestrogenic pesticide methoxychlor (MXC) were evaluated for activity in the presence and absence of Aroclor-1254 induced rat liver S-9 fractions. Both E2 and MXC responded consistently in the assay with average EC₅₀ values of 9.6 x 10^-11 M and 1.2 x 10^-5 M, respectively. In the presence of a 0.1% S-9 fraction, the EC₅₀ for E2 was increased to 1.4 x 10^-9 M and that for MXC decreased to 4.9 x 10^-7 M, with both compounds demonstrating increased secondary metabolite formation as evidenced by HPLC analysis. Consistent with these data, metabolites of E2 and MXC exhibited decreased and increased potencies, respectively, in the assay system relative to the parent molecules. S-9 was compatible with the MCF-7 reporter assay and has the potential to enhance detection of proestrogenic materials. (C) 2000 Elsevier Science Inc.

Original language	English
Pages (from-to)	207-216
Number of pages	10
Journal	Reproductive Toxicology
Volume	14
Issue number	3
DOIs	https://doi.org/10.1016/S0890-6238(00)00070-8
State	Published - May 2000
Externally published	Yes

Keywords

Endocrine toxicity
Estrogenicity
HPTE
MCF-7 cells
Metabolism
Methoxychlor
Reporter gene
S-9 activation

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10.1016/S0890-6238(00)00070-8

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@article{9dc0ad92fb3f466da5086aba10a23a9f,

title = "Incorporation of S-9 activation into an ER-α transactivation assay",

abstract = "We evaluated the feasibility of incorporating an exogenous metabolic activating system into an estrogen receptor-α transactivation assay. 17β- estradiol (E2), and the proestrogenic pesticide methoxychlor (MXC) were evaluated for activity in the presence and absence of Aroclor-1254 induced rat liver S-9 fractions. Both E2 and MXC responded consistently in the assay with average EC50 values of 9.6 x 10-11 M and 1.2 x 10-5 M, respectively. In the presence of a 0.1\% S-9 fraction, the EC50 for E2 was increased to 1.4 x 10-9 M and that for MXC decreased to 4.9 x 10-7 M, with both compounds demonstrating increased secondary metabolite formation as evidenced by HPLC analysis. Consistent with these data, metabolites of E2 and MXC exhibited decreased and increased potencies, respectively, in the assay system relative to the parent molecules. S-9 was compatible with the MCF-7 reporter assay and has the potential to enhance detection of proestrogenic materials. (C) 2000 Elsevier Science Inc.",

keywords = "Endocrine toxicity, Estrogenicity, HPTE, MCF-7 cells, Metabolism, Methoxychlor, Reporter gene, S-9 activation",

author = "Charles, \{Grantley D.\} and Bartels, \{Michael J.\} and C. Gennings and Zacharewski, \{Timothy R.\} and Freshour, \{Nancy L.\} and \{Bhaskar Gollapudi\}, B. and Carney, \{Edward W.\}",

note = "Funding Information: ☆ Support for this research was provided jointly by the Chemical Manufacturers Association (CMA), Reference No. 9010 and The Dow Chemical Company. ",

year = "2000",

month = may,

doi = "10.1016/S0890-6238(00)00070-8",

language = "English",

volume = "14",

pages = "207--216",

journal = "Reproductive Toxicology",

issn = "0890-6238",

publisher = "Elsevier Inc.",

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TY - JOUR

T1 - Incorporation of S-9 activation into an ER-α transactivation assay

AU - Charles, Grantley D.

AU - Bartels, Michael J.

AU - Gennings, C.

AU - Zacharewski, Timothy R.

AU - Freshour, Nancy L.

AU - Bhaskar Gollapudi, B.

AU - Carney, Edward W.

N1 - Funding Information: ☆ Support for this research was provided jointly by the Chemical Manufacturers Association (CMA), Reference No. 9010 and The Dow Chemical Company.

PY - 2000/5

Y1 - 2000/5

N2 - We evaluated the feasibility of incorporating an exogenous metabolic activating system into an estrogen receptor-α transactivation assay. 17β- estradiol (E2), and the proestrogenic pesticide methoxychlor (MXC) were evaluated for activity in the presence and absence of Aroclor-1254 induced rat liver S-9 fractions. Both E2 and MXC responded consistently in the assay with average EC50 values of 9.6 x 10-11 M and 1.2 x 10-5 M, respectively. In the presence of a 0.1% S-9 fraction, the EC50 for E2 was increased to 1.4 x 10-9 M and that for MXC decreased to 4.9 x 10-7 M, with both compounds demonstrating increased secondary metabolite formation as evidenced by HPLC analysis. Consistent with these data, metabolites of E2 and MXC exhibited decreased and increased potencies, respectively, in the assay system relative to the parent molecules. S-9 was compatible with the MCF-7 reporter assay and has the potential to enhance detection of proestrogenic materials. (C) 2000 Elsevier Science Inc.

AB - We evaluated the feasibility of incorporating an exogenous metabolic activating system into an estrogen receptor-α transactivation assay. 17β- estradiol (E2), and the proestrogenic pesticide methoxychlor (MXC) were evaluated for activity in the presence and absence of Aroclor-1254 induced rat liver S-9 fractions. Both E2 and MXC responded consistently in the assay with average EC50 values of 9.6 x 10-11 M and 1.2 x 10-5 M, respectively. In the presence of a 0.1% S-9 fraction, the EC50 for E2 was increased to 1.4 x 10-9 M and that for MXC decreased to 4.9 x 10-7 M, with both compounds demonstrating increased secondary metabolite formation as evidenced by HPLC analysis. Consistent with these data, metabolites of E2 and MXC exhibited decreased and increased potencies, respectively, in the assay system relative to the parent molecules. S-9 was compatible with the MCF-7 reporter assay and has the potential to enhance detection of proestrogenic materials. (C) 2000 Elsevier Science Inc.

KW - Endocrine toxicity

KW - Estrogenicity

KW - HPTE

KW - MCF-7 cells

KW - Metabolism

KW - Methoxychlor

KW - Reporter gene

KW - S-9 activation

UR - https://www.scopus.com/pages/publications/0034045772

U2 - 10.1016/S0890-6238(00)00070-8

DO - 10.1016/S0890-6238(00)00070-8

M3 - Article

C2 - 10838121

AN - SCOPUS:0034045772

SN - 0890-6238

VL - 14

SP - 207

EP - 216

JO - Reproductive Toxicology

JF - Reproductive Toxicology

IS - 3

ER -

Incorporation of S-9 activation into an ER-α transactivation assay

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Keywords

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