ARACHIDONIC ACID REGULATION OF STAR GENE EXPRESSION

花生四烯酸对星基因表达的调控

• 批准号: 6387788
• 负责人: XINGJIA WANG
• 金额: $ 7.4万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-04 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6387788
• 关键词: arachidonate; biological signal transduction; cyclic AMP; gene expression; genetic regulatory element; high performance liquid chromatography; luteinizing hormone; membrane proteins; phosphoproteins; phosphorylation; plasmids; protein kinase A; radioimmunoassay; steroid biosynthesis

The steroidogenic acute regulatory protein (StAR) mediates trophic hormone-stimulated steroid biosynthesis. Since steroid hormones play critical roles in maintaining body homeostasis, the mechanism regulating StAR gene expression is an important component of research on trophic hormone-induced steroidogenesis. It is well known that trophic hormone stimulation induces cyclic AMP (cAMP) formation followed by activation of protein kinase A (PKA) which, through as yet unknown mechanisms, regulate StAR gene expression. However, trophic hormones also induce arachidonic acid release, and inhibition of arachidonic acid release inhibited StAR gene expression and steroidogenesis in spite of high intracellular levels of cAMP. While the mechanism of arachidonic acid regulation of StAR gene expression is not clear, recent studies suggested a scenario in which arachidonic acid and cAMP transduce signals through two separate pathways and co-regulate the transcription factors acting on the StAR promoter. To study this possible mechanism and to prove the co-regulation of these two pathways in StAR gene expression, the following specific Aims are proposed: 1) to demonstrate that in trophic hormone-stimulation, both the arachidonic acid and cAMP-PKA- phosphorylationpathways are required with neither one along being sufficient for StAR gene expression and to study roles of each pathway in steroidogenesis; 2) to determine which arachidonic acid metabolites are involved in trophic hormone-stimulated StAR gene expression and to examine the roles of these metabolites in steroidogenesis; 3) to construct StAR promoter/luciferase reporter plasmids containing serial deletions of the wild type StAR promoter and to test promoter activities with the goal of selecting the region responding to arachidonic acid and identifying the arachidonic acid-response element (ARE) in the selected promoter sequences by continueous deletion and mutagenesis. The resulting ARE DNA sequences will be used in future studies to identify arachidonic acid-regulated transcription factors which may participate in StAR gene transcription. The long term goal of the project is to understand the molecular mechanism responsible for arachidonic acid regulation of hormone-stimulated steroid biosynthesis and the role of this fatty acid in steroid hormone-related physiological activities and diseases including gonadal steroid-related diseases in reproduction.

类固醇激素急性调节蛋白（StAR）介导营养激素刺激的类固醇生物合成。 由于类固醇激素在维持身体体内平衡方面起着关键作用，因此调节星基因表达的机制是营养激素诱导的类固醇发生的研究的重要组成部分。众所周知，营养激素刺激会诱导循环AMP（CAMP）形成，然后激活蛋白激酶A（PKA），通过尚未知道的机制，该蛋白激酶A（PKA）调节了星基因表达。 然而，营养激素还会诱导蛛网膜酸释放，尽管cAMP的细胞内水平很高，但蛛网膜酸释放抑制了恒星基因表达和类固醇生成。 虽然尚不清楚星六烯酸调节星式酸调节的机理，但最近的研究表明，蛛网膜酸和cAMP转导信号通过两种单独的途径，并共同调节作用于星星启动子上的转录因子。 为了研究这种可能的机制并证明这两种途径在星基因表达中的共同调节，提出了以下特定目的：1）证明在营养激素刺激中，蛛网膜酸和cAMP-PKA-PKA-PKA-PKA-PKA-PKA-PHospHoryLationPathways均不需要沿着星基因表达和研究各个路径的绩效，都不足以进行恒星基因表达和研究绩效。 2）确定哪些花生四烯酸代谢产物参与营养激素刺激的恒星基因表达，并检查这些代谢产物在类固醇生成中的作用； 3）构建含有野生型星星启动子的串行缺失的星星/荧光素酶报告基因质粒，并测试启动子活性，目的是选择对蛛网膜酸响应的区域，并通过持续性缺失和烟熏量来识别蛛网膜酸反应元件（IS）。 结果是DNA序列将在以后的研究中使用，以鉴定可能参与Star基因转录的蛛网膜酸调节的转录因子。 该项目的长期目标是了解导致激素刺激的类固醇生物合成的蛛网膜酸调节的分子机制，以及该脂肪酸在类固醇激素相关的生理活性和包括性腺类固醇相关疾病在内的疾病中的作用。

项目成果

Interaction between arachidonic acid and cAMP signaling pathways enhances steroidogenesis and StAR gene expression in MA-10 Leydig tumor cells.

花生四烯酸和 cAMP 信号通路之间的相互作用增强 MA-10 Leydig 肿瘤细胞中的类固醇生成和 StAR 基因表达。

• DOI: 10.1016/s0303-7207(01)00748-1
• 发表时间: 2002
• 期刊: Molecular and cellular endocrinology
• 影响因子: 4.1
• 作者: Wang,XingJia;Dyson,MatthewT;Mondillo,Carolina;Patrignani,Zoraida;Pignataro,Omar;Stocco,DouglasM
• 通讯作者: Stocco,DouglasM

Cyclooxygenase-2 regulation of the age-related decline in testosterone biosynthesis.

• DOI: 10.1210/en.2005-0298
• 发表时间: 2005-10
• 期刊: Endocrinology
• 影响因子: 4.8
• 作者: X. Wang;Chwan-Li Shen;Matthew T. Dyson;S. Eimerl;J. Orly;J. Hutson;D. Stocco

Inhibition of cyclooxygenase-2 activity enhances steroidogenesis and steroidogenic acute regulatory gene expression in MA-10 mouse Leydig cells.

抑制 cyclooxygenase-2 活性可增强 MA-10 小鼠 Leydig 细胞中的类固醇生成和类固醇生成急性调节基因表达。

• DOI: 10.1210/en.2002-0081
• 发表时间: 2003
• 期刊: Endocrinology.
• 作者: Wang,XingJia;Dyson,MatthewT;Jo,Youngah;Stocco,DouglasM
• 通讯作者: Stocco,DouglasM