17 ALPHA HYDROXYLASE EXPRESSION IN HUMAN OVARIAN CELLS

17 人卵巢细胞中的α羟化酶表达

• 批准号: 2357319
• 负责人: Jan M McAllister
• 金额: $ 1.59万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2357319
• 关键词: androgens; cyclic AMP; cytochrome P450; female; gel mobility shift assay; gene expression; gonadotropins; graafian follicles; growth factor; human tissue; messenger RNA; nuclear runoff assay; polycystic ovary syndrome; posttranscriptional RNA processing; protein biosynthesis; protein sequence; steroid 17alpha monooxygenase; steroid hormone biosynthesis; steroid hormone metabolism; tissue /cell culture

DESCRIPTION (Adapted from the Investigator's Abstract): This study will investigate the regulation of human cytochrome P450 17-alpha- hydroxylase (CYP17) gene expression in theca interna cells from ovaries of normal cycling women and from ovaries of women with polycystic ovarian syndrome (PCOS). The hypothesis to be tested is that increased androgen production results from an intrinsic abnormality of steroid production in PCOS theca cells. The goal is to understand how LH and growth factors regulate CYP17 expression and androgen synthesis in normal cells, and how dysregulation of these processes results in increased androgen production in PCOS. Conditions have been developed to propagate functional cultures of normal and PCOS theca cells that express 17a-hydroxylase activity in response to cAMP. In normal, cultured theca cells, cAMP stimulates CYP17 mRNA, whereas epidermal growth factor (EGF), fibroblast growth factor (FGF), and transforming growth factor B (TGFB) inhibit cAMP-stimulation of CYP17 mRNA. The study will characterize the mechanisms by which CYP17 gene expression is stimulated by LH, and inhibited by growth factors in theca cells. Specific Aim 1 will characterize the regulation of CYP17 enzyme activity, protein, and mRNA content in theca cultures, and determine whether the LH-dependent induction of CYP17 mRNA requires ongoing protein synthesis. The investigator will examine the time-and dose-dependent effects of LH and growth factors on CYP17 mRNA levels, and determine whether changes in mRNA stability also contribute to the effects of cAMP and growth factors on steady state CYP17 mRNA. Moreover, the investigator will determine whether these regulatory processes are altered in PCOS. Specific Aim 2 will identify the cis- regulatory elements involved in the tissue-specific regulation of CYP17 gene by LH, the transcription factor steroidogenic factor-1 (SF-1), and growth factors. Should growth factor modulation of CYP17 expression in PCOs theca cells be altered, studies will begin to identify the cis-regulatory elements involved in the regulation of CYP 17 expression by the specific growth factor involved. In Specific Aim 3, studies of steroid metabolism will be performed with fresh explant cultures and long-term cultures of theca cells from normal and PCOS patients to examine whether the abnormalities in the steroidogenic pathway that cause increased androgen production in PCOS are extrinsic or intrinsic. Experiments are planned to determine whether increased androgen production results from changes in the regulation by LH and growth factors of expression of P450 scc (CYP11A), 3B -HSD, CYP17, or some combination of these in PCOS theca cells. Information derived from these studies will provide a better understanding of the molecular basis of steroid synthesis and androgen excess in PCOS.

描述（改编自调查员的摘要）：这项研究将 研究人细胞色素P450 17-α-羟化酶的调节 （CYP17）正常卵巢中theca interna细胞中的基因表达 骑自行车的妇女和多囊卵巢综合征的妇女的卵巢 （PCOS）。 要检验的假设是雄激素产生增加 来自PCOS THECA类固醇产生的固有异常的结果 细胞。 目的是了解LH和增长因素如何调节CYP17 正常细胞中的表达和雄激素合成，以及如何失调 这些过程导致PCOS中的雄激素产生增加。 已经开发了疾病来传播正常的功能培养物 和对17a-羟化酶活性的PCOS theca细胞响应 营。 在正常的，培养的theca细胞中，CAMP刺激CYP17 mRNA，而 表皮生长因子（EGF），成纤维细胞生长因子（FGF）和 转化生长因子B（TGFB）抑制CYP17 mRNA的cAMP刺激。 该研究将表征CYP17基因表达的机制 由LH刺激，并受到theca细胞的生长因子的抑制。 具体的 AIM 1将表征CYP17酶活性，蛋白质的调节 和theca培养物中的mRNA含量，并确定LH依赖性是否依赖 CYP17 mRNA的诱导需要持续的蛋白质合成。 这 研究人员将检查LH和剂量依赖性的效果 CYP17 mRNA水平的生长因子，并确定mRNA的变化是否变化 稳定也有助于营地和成长因素对 稳态CYP17 mRNA。 此外，调查人员将确定是否 这些调节过程在PCOS中发生了变化。 具体目标2将 确定与组织特异性有关的顺式调节元件 通过转录因子类固醇生成的LH调节CYP17基因 因子1（SF-1）和生长因子。 应该调制生长因子 CYP17在PCOS theca细胞中的表达会改变，研究将开始 确定与CYP 17调控有关的顺式调节元素 涉及特定生长因子的表达。 在特定的目标3中 类固醇代谢的研究将使用新鲜的外植体进行 以及正常患者和PCOS患者的theca细胞的长期培养 检查是否引起类固醇途径的异常 PCOS中雄激素产生的增加是外部或固有的。 计划实验以确定雄激素产生是否增加 LH调节的变化和生长因子的变化结果 P450 SCC（CYP11A），3B -HSD，CYP17或某种组合的表达 这些在pcos theca细胞中。 这些研究得出的信息将 更好地理解类固醇合成的分子基础 PCOS中的雄激素过量。