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的蜕膜化 人子宫内膜 孕激素和松弛素（RLX）调节细胞 人的增殖、分化和多基因表达 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）在哺乳动物中广泛分泌， 蜕膜基质细胞 我们假设孕激素依赖性和 RLX诱导的PRL、IGFBP-1和蜕膜特异性核转录 因子调节细胞生长和分化。 目标1： 研究内源性PRL对细胞有丝分裂活性的影响， 人子宫内膜间质细胞DNA合成和瞬时表达 c-fos和c-myc mRNA将在细胞中PRL合成被抑制时检测。 被序列特异性反义寡聚体阻断。 这将揭示 内源性PRL在蜕膜化过程中对细胞增殖的作用。 目的 二是研究非磷酸化抑制作用的机制 和磷酸化IGFBP-1同种型对IGF-1促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 与IGF-I/II结合的IGF-I亚型和IGF-I受体以及相对的 在蜕膜化过程中产生的同种型的量将是 研究了 IGF-I/II对促有丝分裂活性的影响将在 在基质细胞中存在IGFBP-1亚型的情况下进行研究。 目标3是 通过研究基因鉴定蜕膜特异性转录因子 IGFBP-1的转录。实验设计为a）研究 人胰岛素样生长因子结合蛋白-1基因启动子的转录活性 子宫内膜基质细胞和子宫内膜癌细胞系，B）研究 蜕膜核蛋白与远侧和 近端顺式元件，并确定必要的功能顺式元件 对于IGFBP-1基因在蜕膜细胞中的转录，c）鉴定和 描述蜕膜特异性转录的物理特性 因子，d）评估孕酮受体如何影响IGFBP-1 在蜕膜细胞中的转录。这项研究的结果将增加新的 我们对蜕膜细胞基因表达机制的理解 激活和蜕膜化，这是胚泡发育所必需的过程， 着床和维持妊娠。不孕症的影响超过 在美国有200万对夫妇， 流产可能占20%以上。这项研究将进一步促进我们的 了解不孕症的病因，如黄体期 缺陷、着床失败和复发性流产。而且这 该提案可能为开发更安全的避孕方法开辟新的途径 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.