DECIDUAL PROLACTIN IN NORMAL AND PATHOLOGIC PREGNANCIES

相对较少的人知道的分子机制涉及 人类子宫蜕膜化。在过去的几年里，我们实验室 已经开发出一种人类蜕膜化的体外模型系统 提示催乳素是一个模型基因，可以用来描述 参与蜕膜化调控的分子机制。我们 在红豆杉核提取液中发现了几种DNA结合蛋白 未蜕膜的子宫内膜间质细胞在 蜕膜化过程中诱导的几种结合蛋白 蜕膜化。其中两种结合蛋白似乎是蜕膜- 具体的。我们的中心假设是，顺式和顺式 调节蜕膜催乳素表达的调节因子 在蜕膜化过程中将提供对分子的新见解 非女性化进程中涉及的各种机制。我们的具体目标 1)确定蜕膜上顺式作用的DNA元件 催乳素启动子在催乳素基因诱导中的作用 蜕膜化过程中的表达，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，以及3)检验转录 促催乳素基因表达的调节因素 蜕膜化也参与了决定 蜕膜化及其相关基因表达的变化。 将特别强调对新的蜕膜的描述- 特定的转录因子。目标1将使用EMSA、DNase1 缺失和定点定位的足迹和转染法研究 人类原代培养中蜕膜催乳素启动子的突变 子宫内膜细胞。目标2将利用超级换挡和竞争性 结合实验，一个新的转录因子(S)的序列将 在人蜕膜文库的表达克隆后确定。在……里面 目的3.将未蜕膜的子宫内膜间质细胞 过度表达转录因子的表达载体 在目标2中确定，以确定这些因素是否单独和 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的表型变化特征 细胞。总而言之，这些调查应该会提供新的见解 转录级联中涉及的分子机制 催乳素基因表达的相关因素及其调控 人类的蜕膜化和。这样的调查是有实际意义的 重要，因为蜕膜化是成功子宫的关键 种植和蜕膜化异常是常见的原因 人类自然流产的证据。

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.