DECIDUALIZATION OF HUMAN ENDOMETRIAL STROMAL CELLS

本研究的目的是阐明内源性激素和 蜕膜特异性转录因子控制蜕膜化的研究 人子宫内膜。孕激素和松弛素(RLX)对细胞的调节 人类多基因的增殖、分化和表达 子宫内膜间质细胞。催乳素(PRL)和磷酸化胰岛素- 类生长因子结合蛋白-1(IGFBP-1)广泛分泌于 蜕膜间质细胞。我们假设孕激素依赖和 RLX诱导的PRL、IGFBP-1与蜕膜特异性核转录 S因子调节细胞生长和分化。目标1是 探讨内源性催乳素对血管内皮细胞有丝分裂活性的影响 人子宫内膜间质细胞。DNA合成与瞬时表达 当细胞中催乳素合成时，将检测c-fos和c-myc mRNA 被序列特异的反义寡聚物阻断。这将揭示出 蜕膜化过程中内源性催乳素在细胞增殖中的作用目标 二是研究了非磷酸化的抑制作用机理。 和磷酸化的胰岛素样生长因子结合蛋白-L亚型对胰岛素样生长因子-1有丝分裂活性的影响 I/II(胰岛素样生长因子)。IGFBP-1之间的竞争 异构体与胰岛素样生长因子-L受体与胰岛素样生长因子-I/II及其相关分子的结合 蜕膜化过程中产生的异构体数量为 调查过了。IGF-I/II对有丝分裂活性的影响将是 在基质细胞中存在IGFBP-1亚型的情况下进行研究。目标3是 通过对蜕膜特异性转录因子(S)的研究鉴定该基因 IGFBP-1的转录。实验的设计是为了a)研究 人胰岛素样生长因子结合蛋白-1基因启动子转录活性的研究 子宫内膜间质细胞和子宫内膜癌细胞系，b)研究 蜕膜核蛋白与远端蜕膜核蛋白的结合特性 近端顺式元件和鉴定必需的功能顺式元件 对于IGFBP-1基因在蜕膜细胞中的转录，c)鉴定并 描述蜕膜特异性转录的物理特性 因素，d)评估孕酮受体如何影响IGFBP-1 蜕膜细胞中的转录。这项研究的结果将增加新的 我们理解蜕膜细胞基因机制的维度 激活和蜕膜化是胚泡发育所必需的过程 妊娠的植入和维持。不孕不育影响的不仅仅是 美国有200万对夫妇植入失败 流产可能占20%以上。这项研究将进一步推动我们的 了解黄体期等不孕症的病因 缺陷、植入失败和反复妊娠丢失。还有，这个 提案可能为开发更安全的避孕药开辟新途径 方法：研究方法。

The goal of this study is to elucidate the endogenous hormones and decidual-specific transcription factors controlling the decidualization of human endometrium. Progestin and relaxin (RLX) regulate cell proliferation, differentiation and expression of multiple genes in human endometrial stromal cells. Prolactin (PRL) and phosphorylated insulin- like growth factor binding protein-1 (IGFBP-1) are extensively secreted in decidualized stromal cells. We hypothesize that progestin-dependent and RLX-induced PRL, IGFBP-1 and decidual specific nuclear transcription factor(s) regulate cell growth and differentiation. Aim 1 is to investigate the effect of endogenous PRL on the mitogenic activities of human endometrial stromal cells. DNA synthesis and transient expression of c-fos and c-myc mRNAs will be examined when PRL synthesis in cells is blocked by a sequence-specific antisense oligomer. This will reveal the role of endogenous PRL on cell proliferation during decidualization. Aim 2 is to study the mechanism of the inhibitory effect of non-phosphorylated and phosphorylated IGFBP-l isoforms on the mitogenic activities of IGF- I/II (insulin-like growth factor). The competition between IGFBP-1 isoforms and IGF-l receptor for the binding to IGF-I/II and the relative amount of isoforms produced during the process of decidualization will be investigated. The effects of IGF-I/II on the mitogenic activities will be studied in the presence of IGFBP-1 isoform in stromal cells. Aim 3 is to identify decidual-specific transcription factor(s) by studying the gene transcription of IGFBP-1. Experiments are designed to a) study the transcriptional activities of the promoter of IGFBP-1 gene in human endometrial stromal cells and in endometrial cancer cell line, b) study the binding properties between decidual nuclear proteins to the distal and proximal cis-elements and identify the functional cis-elements essential for the transcription of IGFBP-1 gene in decidual cells, c) identify and characterize the physical properties of decidual specific transcription factor, d) evaluate how the progesterone receptor affects the IGFBP-1 transcription in decidual cells. The outcome of this study will add new dimension to our understanding of the mechanism for decidual cell gene activation and decidualization, a process essential for blastocyst implantation and maintenance of pregnancy. Infertility affects more than 2 million couples in the United States in which failure of implantation and miscarriage may comprise more than 20%. This study will further our understanding the causes of infertility diseases such as luteal phase defect, failure in implantation and recurrent pregnancy loss. Also, this proposal may open new avenues for the development of safer contraceptive methods.