ESTROGEN-INDUCED GROWTH FACTOR EXPRESSION IN THE UTERUS

子宫内雌激素诱导的生长因子表达

• 批准号: 3426699
• 负责人: ROBERT D KOOS
• 金额: $ 5万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-01 至 1993-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3426699
• 关键词: RNA directed DNA polymerase; SDS polyacrylamide gel electrophoresis; angiogenesis factor; estrogens; fibroblast growth factor; gene expression; growth factor; hormone regulation /control mechanism; keratinocyte; laboratory rat; messenger RNA; mitogens; nucleic acid hybridization; polymerase chain reaction; uterus; vascular endothelium

The long term goal of this study is to determine how estrogen stimulates growth of the mammalian uterus. Estrogen stimulates hypertrophy and hyperplasia of the uterus and other organs of the female genital tract. This effect is mediated directly at the target tissue via interaction of estrogen with its specific receptor. It is believed that many of estrogen's effects are mediated by growth regulatory factors whose synthesis and/or activity are regulated by estrogen. Despite the significance of these processes to human fertility, relatively little is known about the nature of these factors. In this study, the powerful, sensitive method of reverse transcription-polymerase chain reaction (RT-PCR) will be used to evaluate estrogen's effects on the expression of growth regulatory factors in the rat uterus. In part 1 (Specific Aim I), the ability of estrogen to control the expression of four growth factors that seem likely to play important roles in the cyclic development of the uterus, but whose expression in the uterus is almost completely unexplored, will be determined. These include: 1) acidic fibroblast growth factor (FGF), 2) basic FGF, 3) keratinocyte growth factor (KGF), and 4) vascular endothelial growth factor (VEGF). The FGF's are broad spectrum mitogens that stimulate proliferation of a wide range of cells and are thought to be involved in neovascularization. Expression of basic FGF by human endometrial adenocarcinoma cells has been shown to be stimulated by estrogen. KGF is similar in structure to the FGF's but is specific for epithelial cells; KGF is thought to be a stromally-derived regulator of epithelial cell proliferation. It could, therefore, play a role in the proliferation of endometrial epithelium. VEGF is similar in structure to platelet derived growth factor (PDGF). It is a potent, specific endothelial cell mitogen; in addition it increases vascular permeability. VEGF could play a role in the development and functioning of the uterine vasculature. RNA will be extracted from uteri of control and estrogen-treated immature rats at various intervals after treatment and the expression of the mRNA's for these factors quantitatively analyzed. In part 2 (Specific Aim 2), the induction of as yet undefined mRNA's by estrogen in the uterus will be examined. This will be done using the method of subtraction hybridization followed by PCR amplification of the subtracted mRNA's. To do this, a defined DNA adaptor is ligated to both ends of the products a cDNA synthesis reaction performed on the subtracted mRNA's followed by standard PCR amplification, sequencing of the amplified products, and comparison of the sequences to known genes. In this way, low abundance mRNA's induced by estrogen may be identified; most growth factor mRNA's are relatively rare. These studies will provide new insight into the nature of the growth regulating factors involved in sex steroid hormone action on the female genital tract.

这项研究的长期目标是确定雌激素如何刺激 哺乳动物子宫的生长。雌激素刺激肥大和 子宫的增生和女性生殖道的其他器官。 这种作用是通过相互作用直接在目标组织介导的 雌激素具有特定受体。 相信许多 雌激素的作用是由生长调节因素介导的 合成和/或活性受雌激素调节。 尽管有 这些过程对人类生育的重要性，相对较少的是 知道这些因素的性质。 在这项研究中，强大的 逆转录聚合酶链反应的敏感方法 （RT-PCR）将用于评估雌激素对表达的影响 大鼠子宫的生长调节因子。 在第1部分（特定目标I）， 雌激素控制四个生长因子的表达的能力 这似乎在循环发展中起着重要作用 子宫，但在子宫中的表达几乎完全没有探索， 将确定。其中包括：1）酸性成纤维细胞生长因子 （FGF），2）基本FGF，3）角质细胞生长因子（kgf）和4）血管 内皮生长因子（VEGF）。 FGF是广泛的有丝分裂剂 刺激广泛细胞的增殖，被认为是 参与新血管形成。 人类表达基本fgf 子宫内膜腺癌细胞已被证明是由 雌激素。 kgf的结构与FGF相似，但特定于 上皮细胞； KGF被认为是基本的调节器 上皮细胞增殖。 因此，它可以在 子宫内膜上皮的增殖。 VEGF的结构与 血小板得出的生长因子（PDGF）。这是一个有效的特定内皮 细胞丝裂原;另外，它增加了血管渗透性。 VEGF可以 在子宫脉管系统的发展和功能中发挥作用。 RNA将从对照和雌激素治疗的未成熟的子宫中提取 治疗后的各个间隔和mRNA的表达 对于这些因素，已定量分析。 在第2部分（特定目标2）中， 雌激素在子宫中诱导AS尚未定义的mRNA是 检查。 这将使用减法杂交方法完成 然后进行PCR放大减法mRNA。 为此， 定义的DNA适配器连接到产品的两端A cDNA 在减去mRNA的综合反应，然后是标准 PCR扩增，放大产物的测序以及比较 已知基因的序列。 这样，低丰度mRNA的诱导 可以鉴定出雌激素；大多数生长因子mRNA相对较少。 这些研究将为增长的本质提供新的见解 调节性类固醇激素作用对女性的调节因素 生殖道。