DECIDUAL PROLACTIN IN NORMAL AND PATHOLOGIC PREGNANCIES

相对较少的是所知的分子机制， 人类子宫蜕膜化 在过去的几年里，我们的实验室 已经开发了一种体外人类蜕膜化模型系统， 表明催乳素是一个模型基因， 参与调控蜕膜化的分子机制。 我们 已经鉴定了几种DNA结合蛋白在核提取物中， 未蜕膜化的子宫内膜间质细胞在子宫内膜异位症中表达下调， 蜕膜化和几种结合蛋白的诱导过程中， 蜕膜化 其中两种结合蛋白似乎是蜕膜- 特定. 我们的中心假设是顺式和 调节蜕膜催乳素表达的反式调节因子 在蜕膜化过程中将提供新的见解， 参与蜕膜化过程的机制。 我们的具体目标 是1）确定蜕膜上的顺式作用DNA元件 催乳素启动子对催乳素基因诱导至关重要 在蜕膜化过程中表达，2）识别和表征 介导催乳素基因诱导的转录因子 在蜕膜化过程中，和3）测试的假设，转录 促乳素基因表达的诱导因子 蜕膜化也参与决定 蜕膜化和相关的基因表达变化。 特别强调将给予新的蜕膜表征- 特异性转录因子Aim 1将使用EMSA、DNase 1 足迹法和转染研究， 原代培养人蜕膜催乳素启动子的突变体 子宫内膜细胞 目标2将利用超移和竞争力 结合测定，并且新转录因子的序列将 在人蜕膜文库的表达克隆后确定。 在 目的3、转染未蜕膜化子宫内膜间质细胞， 过表达转录因子的表达质粒 目标2中确定的，以确定这些因素是否单独存在， 结合，诱导细胞表达催乳素并获得其他 蜕膜化子宫内膜间质表型特征性变化 细胞 综合起来，这些调查应该会提供新的见解 参与转录级联反应的分子机制 参与催乳素基因表达的因素和 人类蜕膜化和。这些研究具有一定的实用价值 重要性，因为蜕膜化对于成功子宫 着床和蜕膜化异常是常见的原因 人类自然流产

Relatively little is known about the molecular mechanisms involved in human uterine decidualization. Over the past few years, our laboratory has developed an in vitro model system of human decidualization that indicates that prolactin is a model gene with which to delineate the molecular mechanisms involved in the regulation of decidualization. We have identified several DNA binding proteins in nuclear extracts of undecidualized endometrial stromal cells that are downregulated during decidualization and several binding proteins that are induced during decidualization. Two of these binding proteins appear to be decidua- specific. Our central hypothesis is that delineation of the cis- and transacting factors regulating the expression of decidual prolactin during decidualization will provide new insights into the molecular mechanisms involved in the decidualization process. Our specific aims are to 1) determine the cis-acting DNA elements on the decidual prolactin promoter that are critical for induction of prolactin gene expression during decidualization, 2) identify and characterize the transcription factors that mediate induction of the prolactin gene during decidualization, and 3) test the hypothesis that transcription factors that mediate induction of prolactin gene expression during decidualization are also involved in the determination of decidualization and the associated changes in gene expression. Particular emphasis will be given to characterization of novel decidua- specific transcription factors. Aim 1 will utilize EMSAs, DNase 1 footprinting and transfection studies with deletion and site-directed mutants of the decidual prolactin promoter in primary cultures of human endometrial cells. Aim 2 will utilize supershift and competitive binding assays, and the sequence of a novel transcription factor(s) will be determined after expression cloning of a human decidual library. In Aim 3, undecidualized endometrial stromal cells will be transfected with expression plasmids that overexpress the transcription factors identified in Aim 2 to determine whether these factors, alone and in combination, induce the cells to express prolactin and acquire other phenotypic changes characteristic of decidualized endometrial stromal cells. Taken together, these investigations should provide new insights into the molecular mechanisms involved in the cascade of transcription factors involved in prolactin gene expression and the regulation of human decidualization and. Such investigations are of practical importance since decidualization is essential for successful uterine implantation and abnormalities of decidualization are frequent causes of spontaneous human abortion.