Estrogen binding capacity in cytosol fraction of human uterine cervix: effect of estrogen and anti-estrogens

Shunzo Kobayashi
In: Okayama Igakkai Zasshi (Journal of Okayama Medical Association) · 1992 · vol. 104(1-2) , pp. 137–144 · doi:10.4044/joma1947.104.1-2_137 · W2513052400
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This study measured estrogen binding activity in human uterine cervix cytosol and found that synthetic estrogens, anti-estrogens, androgens, gestagens, and danazol inhibited binding non-competitively.

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The study measured estrogen binding activity in the cytosol fraction of human uterine cervix tissue by incubating samples with 10 nM [3H] estradiol-17β, separating bound from free hormone using dextran-coated charcoal, and deriving binding parameters with a Scatchard plot (reported Kd = 2.0×10−9 M). A range of steroids were tested at 1.0 μM to assess their effects on estrogen binding, with natural and synthesized steroids showing inhibitory impacts in different strength categories: synthetic estrogens strongly inhibited, anti-estrogens strongly to moderately inhibited, androgens moderately inhibited, and gestagens moderately inhibited. Danazol was found to inhibit estrogen binding to an extent comparable to androgens, and all observed inhibition was described as non-competitive. This paper is relevant to endometriosis because it specifically reports that danazol—used for the treatment of endometriosis—was as effective as androgens in inhibiting estrogen binding in cervical cytosol.

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Abstract

The estrogen binding activity in the human uterine cervix was measured, and the effect of natural and synthesized steroids on the activities was determined. To assay the estrogen binding activity, the sample was incubated with 10 nM [3H] estradiol-17β at 30°C for 2 hours. Then dextran-coated charcoal (DCC) was added to a final concentration of 0.5% to separate bound/free (B/F) estradiol. Estrogen binding activity was determined by subtracting the activity found in the heated sample from the corresponding activity in the untreated sample. For this purpose it was found appropriate to heat the sample at 40°C for 60 minutes. The dissociation constant obtained from the Scatchard plot analysis was: Kd=2.0×10-9M. A variety of steroids at the same molar concentration (1.0μM) were added to the sample to determine their effects on the estrogen binding activity. For binding with [3H] estradiol-17β, the synthetic estrogens were strongly inhibitory, the anti-estrogen agents were strongly to moderately inhibitory, the androdgens were moderately inhibitory, and the gestagens were moderately inhibitory. Danazol, which has been used for the treatment of endometriosis, was found to be as effective as androgens. All of those inhibitory effects occurred in a non-competitive manner.
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ヒト子宮頸部組織における細胞質分画エストロゲン結合能とそれに影響する因子について 1992 年 104 巻 1-2 号 p. 137-144 詳細 抄録 The estrogen binding activity in the human uterine cervix was measured, and the effect of natural and synthesized steroids on the activities was determined. To assay the estrogen binding activity, the sample was incubated with 10 nM [3H] estradiol-17β at 30°C for 2 hours. Then dextran-coated charcoal (DCC) was added to a final concentration of 0.5% to separate bound/free (B/F) estradiol. Estrogen binding activity was determined by subtracting the activity found in the heated sample from the corresponding activity in the untreated sample. For this purpose it was found appropriate to heat the sample at 40°C for 60 minutes. The dissociation constant obtained from the Scatchard plot analysis was: Kd=2.0×10-9M. A variety of steroids at the same molar concentration (1.0μM) were added to the sample to determine their effects on the estrogen binding activity. For binding with [3H] estradiol-17β, the synthetic estrogens were strongly inhibitory, the anti-estrogen agents were strongly to moderately inhibitory, the androdgens were moderately inhibitory, and the gestagens were moderately inhibitory. Danazol, which has been used for the treatment of endometriosis, was found to be as effective as androgens. All of those inhibitory effects occurred in a non-competitive manner. © 岡山医学会

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