Androgen Receptor in Serially Subcultured Human Endometrial Fibroblasts

Abstract

Specific dihydrotestosterone (DHT) binding was demonstrated in the whole cell sonicates obtained from serially subcultured human endometrial fibroblasts. This binding showed a high affinity and specificity for DHT (Kd=3.3×10^-10M). Various kinds of excess nonradioactive steroids were arranged, as a result of competition among them, in the following decreasing order of binding affinity: DHT, testosterone, cyproterone acetate, 17β-estradiol, progesterone, and 5α-androstane-3α, 17β-diol. Androsterone, 5α-androstanedione, cortisol, androstenedione and estrone hardly affected the binding even at 200 fold excess. Saturation analysis indicated that cultured endometrial and genital skin fibroblasts have a similar number of binding sites for DHT (14.3±8.5 vs 11.5±8.6 fmoles/mg cell protein). Sucrose density gradient centrifugation brought about a sharp peak of ³H-DHT in the 4S region in high ionic strength buffer. This binding was abolished by heating sonicates at 37°C for 1 hr, treating them with pronase or adding to them excess nonradioactive DHT. Endometrial fibroblasts also contained testosterone binding components which formed a peak in the 4S region and showed a similar binding affinity to that for DHT.