Kaufman M, Pinsky L, Hollander R, Bailey J D
J Steroid Biochem. 1983 Apr;18(4):383-90. doi: 10.1016/0022-4731(83)90055-9.
Normal genital skin fibroblast (GSF) monolayers incubated with serum-free medium containing 3 nM [3H]-5 alpha-dihydrotestosterone (DHT) at 37 degrees C for 20 h have about 35% more specific DHT-binding than replicates incubated in serum-free medium with [3H]-DHT for only 1 h to saturate basal specific androgen-receptor activity. If, after 19 h, spent medium is replaced by fresh medium with 3 nM [3H]-DHT for 1 h, specific DHT binding is 85% more than basal. The acquisition of increased binding is temperature dependent (37 greater than 27 degrees C) and cycloheximide (2 microM) suppressible. The increased binding activity is considered to represent an augmentation of androgen receptor concentration because it has the same equilibrium dissociation constant (KD approximately 0.5 nM), rate constant of dissociation (k-1 approximately 6 x 10(-3) min-1) and ligand specificity as basal androgen-receptor activity, and because basal DHT-binding activity is stable in cells preincubated in androgen-free or serum-free medium alone for up to 72 h before assay. Prolonged incubation with methyltrienolone (R1881), a nonmetabolizable synthetic androgen, causes a greater, more persistent increment of androgen receptor activity than does equimolar DHT. The fibroblasts from two subjects with receptor-positive, partial androgen resistance lose their basal receptor activity during prolonged incubation with DHT, but augment it normally with R1881. This suggests that defective DHT metabolism is somehow involved in the pathogenesis of their androgen resistance.
正常生殖器皮肤成纤维细胞（GSF）单层在37℃下与含有3 nM [3H]-5α-二氢睾酮（DHT）的无血清培养基孵育20小时，其特异性DHT结合比在无血清培养基中仅与[3H]-DHT孵育1小时以饱和基础特异性雄激素受体活性的重复样本多约35%。如果在19小时后，将用过的培养基换成含有3 nM [3H]-DHT的新鲜培养基再孵育1小时，特异性DHT结合比基础水平高85%。增加的结合的获得是温度依赖性的（37℃大于27℃）且可被环己酰亚胺（2μM）抑制。增加的结合活性被认为代表雄激素受体浓度的增加，因为它具有与基础雄激素受体活性相同的平衡解离常数（KD约为0.5 nM）、解离速率常数（k-1约为6×10-3分钟-1）和配体特异性，并且因为基础DHT结合活性在仅在无雄激素或无血清培养基中预孵育长达72小时然后进行测定的细胞中是稳定的。与不可代谢的合成雄激素甲基三烯醇酮（R1881）长时间孵育比等摩尔的DHT导致更大、更持久的雄激素受体活性增加。两名受体阳性、部分雄激素抵抗患者的成纤维细胞在与DHT长时间孵育期间失去其基础受体活性，但用R1881能正常增加其活性。这表明有缺陷的DHT代谢以某种方式参与了他们雄激素抵抗的发病机制。
