REGULATION OF HUMAN ESTROGEN SYNTHETASE

人体雌激素生物合成调控的研究进展 类固醇激素生物化学中几乎没有探索过的前沿领域之一 这对生殖等不同领域产生了重要影响， 性发育和癌症。这项研究中提出的实验是 旨在研究生物活性和生物合成的机制 雌激素合成酶(芳香酶)，一种将雄激素转化为 雌激素，在类固醇合成组织中受自然过程调节。 人足月胎盘组织微体部分的芳香酶 丰富和容易获得的人类酶来源，将被提纯和 细胞色素P-450单加氧酶系统的组成 将对芳构化进行鉴定和表征。这个 从其纯化的组分中重建芳香酶活性将是 以及环境的影响(例如，pH，离子强度， 温度、脂质成分)、类固醇(例如孕激素、皮质激素， 雄激素、雌激素)和非类固醇化合物(例如，促性腺激素， 重组芳香酶和胎盘上的前列腺素 微体芳香酶。芳香酸酶的限速成分 人胎盘微粒体、滋养层细胞培养 绒毛膜癌和人卵巢组织匀浆将通过 将每个纯化的芳香酶组分单独加入到 感兴趣系统的膜分数并测量其对 芳构化率。纯化的芳香酶组份的抗体将 获得并使用，与放射性脉冲标记一起使用 氨基酸，以测量这些成分在体内的生物合成速度 类固醇生成组织(原代胎盘细胞/器官培养，绒毛膜癌 细胞培养、原代卵巢颗粒细胞培养)前后 促性腺激素、前列腺素和雌激素刺激雌激素分泌 环状AMP。

Investigation into the regulation of estrogen biosynthesis in the human is one of the virtually unexplores frontiers of steroid hormone biochemistry that has important ramifications into such diverse areas as reproduction, sexual development and cancer. The experiments proposed in this study are designed to investigate mechanisms by which the activity and biosynthesis of estrogen synthetase (aromatase), the enzyme which converts androgens to estrogens, are regulated by natural processes in steroidogenic tissue. Aromatase from the microsomal fraction of human term placental tissue, a rich and easily available source of the human enzyme, will be purified and the components of the cytochrome P-450 mono-oxygenase system responsible for aromatization will be identified and characterized. The re-constitution of aromatase activity from its purified components will be studied as well as the effects of environment (e.g., pH, ionic strength, temperature, lipid composition), steroid (e.g., progestins, corticoids, androgens, estrogens) and non-steroid compounds (e.g., gonadotropins, protaglandins, cyclic AMP) on reconstituted aromatase and on placental microsomal aromatase. The rate-limitine component of aromatase in placental microsomes, trophoblast cell culture derived from human choriocarcinoma, and human ovarian tissued homogenate will be determined by incorporating each purified aromatase component individually into the membrane fraction of the system of interest and measuring the effect on the aromatization rate. Antibody to the purified aromatase components will be obtained and used, in conjunction with pulse-labeling with radioactive amino acids, to measure the rate of biosynthesis of these components in steroidogenic tissue (primary placental cell/organ culture, choriocarcinoma cell culture, primary ovarian granulosa cell culture) before and after stimulation of estrogen secretion by gonadotropins, prostaglandins and cyclic AMP.