Neubauer B L, Anderson N G, Cunha G R, Towell J F, Chung L W
J Steroid Biochem. 1985 Jul;23(1):95-101. doi: 10.1016/0022-4731(85)90266-3.
Epithelium of the adult mouse urinary bladder (BLE) was experimentally combined with mesenchyme of the urogenital sinus (UGM) and grown in intact male hosts to produce prostate-like glandular structures. To determine the extent to which the BLE is altered in a functional sense by inductive influences from UGM, investigations into the in vitro metabolism of tritiated testosterone (T) were undertaken. An isocratic high performance liquid chromatographic (HPLC) method was developed in order to separate the metabolites of T in mouse bladder, prostate and UGM + BLE tissue recombinants. Using a C-18 reversed phase column and a tetrahydrofuran (20): methanol (40): H2O (40) mobile phase, efficient and rapid separation of T, dihydrotestosterone, 3 alpha-androstanediol, androstenedione, androstanedione and androsterone was achieved. The identities of the radiolabeled T metabolites were confirmed by recrystallization to constant specific activity. The results of the present study revealed that tissue recombinants expressed testosterone metabolic profiles only partially toward that of the adult prostate. For example, percentage formation of 5 alpha-androstanedione, 3 alpha-androstanediol and unknown polar metabolites in the UGM + BLE resembled the prostate and differed significantly from the urinary bladder. Conversely, formation of the 3 beta-androstanediol and androsterone from testosterone resembled the urinary bladder and differed from the formation of these metabolites in the prostate. These results suggest that in contrast to histomorphology, androgen-induced DNA synthesis, androgen receptor binding activity and total tissue two-dimensional gel electrophoretic protein profiles, androgen metabolic profiles in the tissue recombinants showed only partial transformation into prostatic phenotypes. Analysis of steroid-metabolic profiles, therefore, may represent an exquisite and sensitive method to assess gene expression in various hormone-responsive target tissues.
将成年小鼠膀胱（BLE）上皮与泌尿生殖窦（UGM）间充质进行实验性组合，并在完整雄性宿主中培养，以生成前列腺样腺结构。为了确定BLE在功能上受UGM诱导影响而改变的程度，对氚标记睾酮（T）的体外代谢进行了研究。开发了一种等度高效液相色谱（HPLC）方法，以分离小鼠膀胱、前列腺以及UGM + BLE组织重组体中T的代谢物。使用C - 18反相柱和四氢呋喃（20）：甲醇（40）：水（40）流动相，实现了T、二氢睾酮、3α - 雄烷二醇、雄烯二酮、5α - 雄烷二酮和雄酮的高效快速分离。通过重结晶至恒定比活度来确认放射性标记T代谢物的身份。本研究结果表明，组织重组体仅部分表达出成年前列腺的睾酮代谢谱。例如，UGM + BLE中5α - 雄烷二酮、3α - 雄烷二醇和未知极性代谢物的生成百分比与前列腺相似，与膀胱有显著差异。相反，睾酮生成3β - 雄烷二醇和雄酮的情况与膀胱相似，与前列腺中这些代谢物的生成情况不同。这些结果表明，与组织形态学、雄激素诱导的DNA合成、雄激素受体结合活性以及全组织二维凝胶电泳蛋白质谱不同，组织重组体中的雄激素代谢谱仅部分转化为前列腺表型。因此，类固醇代谢谱分析可能是评估各种激素反应性靶组织中基因表达的一种精确且灵敏的方法。
