Environmental Toxins and Uterine Gene Expression

环境毒素和子宫基因表达

基本信息

批准号：
7891286
负责人：
SANJOY K. DAS
金额：
$ 33.41万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-05-01 至 2013-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7891286
关键词：
Address Animal Model Biological Biological Models Biology CCL4 gene Cell Nucleolus Cell Nucleus Cell Proliferation Cells Chromatin Coculture Techniques Complement Complex DNA biosynthesis Endoplasmic Reticulum Epithelial Cell Proliferation Estradiol Estrogen Antagonists Estrogen Nuclear Receptor Estrogen Receptors Estrogens Event Figs - dietary G-Protein-Coupled Receptors Gene Expression Gene Expression Regulation Genes Genetic Transcription Goals Growth Growth Factor ICI 182780 In Vitro Injection of therapeutic agent Intervention Investigation Knockout Mice Lactoferrin Late Effects Ligands Mediating Membrane Modeling Molecular Molecular Genetics Mus Organ Phase Phosphorylation Physiological Play Positioning Attribute Protein Biosynthesis Protein Isoforms Proteins Recruitment Activity Regulation Role Signal Pathway Signal Transduction Stimulus Stream System Testing Tissues Toxic Environmental Substances Uterus Water Weight Wild Type Mouse Work cell growth in vitro Model in vivo inhibitor/antagonist insight kepone knock-down non-genomic novel promoter public health relevance response uptake xenoestrogen

项目摘要

DESCRIPTION (provided by applicant): Early- and late-phase estrogenic responses in the uterus have been recognized for more than 60 years, yet mechanisms involved in their regulation remain controversial. One concept is that an early events(s), occurring within the first 6 h, prepares the uterus for later (18-30 h) increase in DNA synthesis, cell proliferation and protein synthesis. An alternate view is that the late growth phase is a result of the continuous presence of a stimulus. Discussion of either concept usually makes the assumption that all of the responses are dependent upon ligand interaction with one of the two estrogen receptor isoforms (ER1 and ER2). However, increased gene expression following injection of estrogen or xenoestrogen, in mice lacking ER1, or in which all ER-activity has been suppressed by an estrogen-antagonist, ICI 182,780 (ICI), has shown this to be an oversimplification. In this regard, accumulating evidence suggests that estrogen regulates diverse but interdependent signaling pathways in uterine biology via ER- dependent and -independent manners. While our long standing hypothesis is that estrogenic certain early responses are ER-independent, late responses are ER1- dependent, and a cross-talk between the two phases is necessary for a full complement of estrogenic responses in the uterus. Our previous and preliminary observations suggest that among several such early genes, Bip and Sik-SP are induced by estradiol- 172 (E2) and kepone in the mouse uterus via ER-independent manner as a phase-I response. Moreover, we recently showed that Bip is critically necessary in mediating E2- or kepone-dependent estrogen signaling that involves ER1 functions. Our preliminary studies also indicated that E2 induces GPR30 and ERK1/2 phoshorylation without involving ER1. Furthermore, we observed that the administration of a selective inhibitor of ERK1/2 activation (SL327), 30 min prior to E2 injection abrogates early ER- independent genes, as well as uterine wet weights during the early and late responsive phases. In contrast, injection of the inhibitor 6 h after E2 injection did not alter late effects. Collectively, these results positioned us to study early and late estrogenic responses to determine molecular relationship between the phases. Our specific aims are to elucidate: 1. Early molecular signaling by E2 or kepone involving Sik-SP, Bip and GPR30 in the uterus. 2. Functional significance of Sik-SP and GPR30 in E2- or kepone- mediated late estrogenic responses in the uterus. Studies should yield new molecular insights involving Bip, Sik-SP and GPR30 during ER1-mediated estrogen signaling. Overall, the results will help us to define relationship and a molecular cross-talk between the two-phase estrogenic responses in the uterus. PUBLIC HEALTH RELEVANCE: Our specific aims are to elucidate: 1. Early molecular signaling by E2 or kepone involving Sik-SP, Bip and GPR30 in the uterus. 2. Functional significance of Sik-SP and GPR30 in E2 or kepone mediated late estrogenic responses in the uterus. Studies should yield new molecular insights involving Bip, Sik-SP and GPR30 regulation of ER1-mediated estrogen signaling. Overall, the results will help us to define the relationship and a molecular cross-talk between the two-phase estrogenic responses in the uterus.

描述（由申请人提供）： 60 多年来，人们已经认识到子宫的早期和晚期雌激素反应，但其调节机制仍然存在争议。一个概念是，在最初 6 小时内发生的早期事件，为子宫随后（18-30 小时）DNA 合成、细胞增殖和蛋白质合成的增加做好了准备。另一种观点认为，生长后期是持续存在刺激的结果。对任一概念的讨论通常假设所有反应都依赖于配体与两种雌激素受体亚型（ER1 和 ER2）之一的相互作用。然而，在缺乏 ER1 或所有 ER 活性均被雌激素拮抗剂 ICI 182,780 (ICI) 抑制的小鼠中，注射雌激素或异雌激素后基因表达增加，表明这种情况过于简单化。在这方面，越来越多的证据表明，雌激素通过 ER 依赖性和非依赖性方式调节子宫生物学中多种但相互依赖的信号通路。虽然我们长期以来的假设是雌激素的某些早期反应与 ER 无关，但晚期反应与 ER1 相关，并且两个阶段之间的串扰对于子宫中雌激素反应的全面补充是必要的。我们之前和初步的观察表明，在几个这样的早期基因中，Bip 和 Sik-SP 是由小鼠子宫中的雌二醇-172 (E2) 和开酮通过 ER 独立的方式诱导的，作为 I 期反应。此外，我们最近表明，Bip 在介导涉及 ER1 功能的 E2 或酮依赖性雌激素信号传导中至关重要。我们的初步研究还表明，E2 诱导 GPR30 和 ERK1/2 磷酸化，而不涉及 ER1。此外，我们观察到，在 E2 注射前 30 分钟施用选择性 ERK1/2 激活抑制剂 (SL327) 会消除早期 ER 独立基因，以及早期和晚期反应阶段的子宫湿重。相反，注射 E2 后 6 小时注射抑制剂并没有改变后期效应。总的来说，这些结果使我们能够研究早期和晚期雌激素反应，以确定各阶段之间的分子关系。我们的具体目标是阐明： 1. E2 或 kepone 涉及子宫中的 Sik-SP、Bip 和 GPR30 的早期分子信号传导。 2. Sik-SP 和 GPR30 在 E2 或开酮介导的子宫晚期雌激素反应中的功能意义。研究应该会在 ER1 介导的雌激素信号传导过程中产生涉及 Bip、Sik-SP 和 GPR30 的新分子见解。总的来说，这些结果将帮助我们定义子宫内两相雌激素反应之间的关系和分子串扰。公共健康相关性：我们的具体目标是阐明： 1. E2 或 kepone 涉及子宫内 Sik-SP、Bip 和 GPR30 的早期分子信号传导。 2. Sik-SP和GPR30在E2或kepone介导的子宫晚期雌激素反应中的功能意义。研究应该会产生新的分子见解，涉及 Bip、Sik-SP 和 GPR30 对 ER1 介导的雌激素信号传导的调节。总的来说，这些结果将帮助我们定义子宫内两相雌激素反应之间的关系和分子串扰。