Study on the cytosol and nuclear androgen receptor of normal male adults and patients with androgen insensitivity syndrome
CUI YuGui,ZHANG GuiYuan,LI Yian,YU Qi,He Fangfang,TIAN Qinjie,GE Qinsheng De partment of Reproductice Endocrinology,National Research Institute for Family Planning, Beijing 100081 *Peking Union Medical College Hospilal
bjectives:To develop a method for measuring directly the cytosol and nuclear androgen receptor(AR)without tissue and cell culture.Design:A laboratory methodology and its clinical application. Setting:The Reproductive Endoerinology Laboratory of National Research Institute for Family Plan-ning. Suhjects:11 Chinese cdult men,3 patients with complete androgen insensitive Syndrome(CAIS)and 6 Intervention:Foreskin specimen were taken from the volunteer at the time of circumcision, the genital skin of patient were biopsied with his consent.Tissues were quickly frozen in liquid nitrogen and stored at -70℃. The fresh rat liver tissue’s were prepared at the day of receptor assay. Methods:About 0.5～1.0g tissues were homogenized directly and centrifuged at 12000 g to get cy-tosol.The nucleochylema were extracted with 0.6 mol/L KCl buffer from the nuclear pellets.Total protein was measured by Bradford’s method.Triamcinolone acetonide was used to reduce non-specific binding. The receptor radioassay(RRA ) was in the principle of‘Scatchard Plots’and calculated with the computer pro-gramme‘Symphony Release 2.0’.Results:The cytosol AR was chracterized by Bm 24.03± 7.19(x±s) fmol/mg protein and Kd 15.13×1O-10mol/L,and nuclear AR,Bm204.37±119. 62 mol/mg protein,Kd 6.38× 1O-10mol/L The capaci-ties of cytosol AR of 3 patients with CAIS were 2.29,O. 60 and 1. 35 fmol/mg protein;nuclear AR,13.22,7. 28 and 6.11 fmol/mg protein. The cytosol of rat livers, Bm 4. 43～5. 25 fmol/mg protein and Kd 7. 29～13. 80× 10-10mol/L,were good quality control. Conclusion:The method,without tissue and fibroblast culture, was more practical and easy to per-fom. The nuclear AR was of much higher level and higher affinity to ligand than the eytosol AR.The vol-umes of AR(espicially nuclear AR)of 3 patients with CAIS were much lower than those of normal male adults. It shown that these androgen resistances were caused by AR defect.
【CateGory Index】： R697