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 EC 50 values of 9.6 × 10 −11 M and 1.2 × 10 −5 M, respectively. In the presence of a 0.1% S-9 fraction, the EC 50 for E2 was increased to 1.4 × 10 −9 M and that for MXC decreased to 4.9 × 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.