男性外阴部皮肤成纤维细胞雄激素受体的研究──（二）核特异结合及受体温度稳定性的检测

STUDIES OF ANDROGEN RECEPTORS IN GENITAL SKIN FIBROBLASTS: ASSAYS OF NUCLEAR SPECIFIC BINDING OF ￣3H-R1881 AND TEMPERATURE STABILITY OF ANDROGEN RECEPTORS

在完整细胞与３Ｈ－Ｒ１８８１特异结合的基础上，用ＳＴＭ－ＴｒｉｔｏｎＸ－１００缓冲液分离粗制核，建立了简便而快速地检测核雄激素特异结合的方法。对正常男性包皮成纤维细胞核与３Ｈ－Ｒ１８８１特异结合的Ｓｃａｔｃｈａｒｄ及单点分析表明，该细胞核与３Ｈ－Ｒ１８８１的特异结合具有高亲和力、低容量的特征，当细胞与近饱和浓度的３Ｈ－Ｒ１８８１温育５０ｍｉｎ后，４７．１６±７．９９％（ｎ＝２０）的特异结合被发现存在于核内，而一睾丸女性化（ｔｆｍ）综合征患者的核特异结合量明显低于该值。我们还显示，正常男性包皮成纤维细胞与３Ｈ—Ｒ１８８１在４２℃温育时的特异结合量比在３７℃温育时减少，但其量低于４０％。当减少量超过４０％时，表明存在热不稳定型的雄激素受体。

By using a simple method, the presence of saturable and high affinity specific binding sits of 3H-R1881 in isolated crude nuclei of cultured genital skin fibroblsts were revealed.When intact foreskin fibroblasts from 20 normal men were incubated at 37 C with saturating concentration (2. 4 nmol/L ) of 3H-R1881, 47. 16±7. 99% of the specific binding was recovered in crude nuclei. Temperature stability of androgen receptors (AR) in fibroblasts was also tested.Specific binding of 3H- R1881 in these cells from normal men decreased by less than 40% when the incubating temperature is raised to 42℃. A greater than 40% decrease of specific binding at 42℃ compared to the value at 37℃ was considered evidence for thermolability.