Developmental and reproductive function of estrogen signaling

雌激素信号的发育和生殖功能

• 批准号: 13671162
• 负责人: TSUKUI Tohru
• 金额: $ 2.24万
• 依托单位: Saitama Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671162/
• 关键词: Estrogen signaling; Estrogen responsive genes; Cell proliferation; Conditional transgenic; Mammary tumor; Bone metabolism; エストロゲンシグル; エストロゲシ応答遺伝子; コンディショナルコンディショナルトランスジェニック; エストロゲンシグナル伝達; 発生工学; 発生生物学; 生殖生理学; トランスジェニックマウス; 組換えアデノウイルス; Wntシグナ

Estrogen and its receptors (ERα and ERβ) are not only essential in female for normal function, but also necessary in male function such as reproductive tract, sexual behavior, maintenance of the skeleton, and cardiovascular system. In general, physiological estrogen action is regulated the transcription of estrogen responsive genes mediated by estrogen receptors. So far, the mechanism of specific estrogen action is still not clear. Simple question is what is estrogen action, and how to make specific estrogen action in vivo.In this experiment, we have focused on gain of function in mouse in vivo by using recombinant adenovirus vector and/or conditional transgenic mice, which expressed constitutive active form human ERα and ERβ, such as ca ERα and ERβ, respectively. Alternative strategy was "conditional gain of function" by using transgenic mice with Cre/loxP system. These transgenics were expressed the reporter genes (nlsGFP/DsRed2) before treated wo/Cre recombinase, were expressed the caERα and caERβ after treated w/Cre, respectively. It was possible to activate the estrogen signaling in the timing and organs selectively, treated with Cre recombinase.Finally, our goal is unveiled with the physiological role of estrogen signaling by through "selective gain/loss of function" in vivo. In this projects, we show that efp (estrogen responsive finger protein) is required for cell proliferation in mammary tumor by through the proteolysis 14-3-3 sigma which is negative regulator cell cycle (Nature, 417, 871-875, 2002).

雌激素及其受体(ER-α和ER-β)不仅是女性正常功能所必需的，也是男性生殖道、性行为、骨骼维护和心血管系统等功能所必需的。一般来说，生理雌激素的作用是由雌激素受体介导的雌激素反应基因的转录调节的。到目前为止，雌激素的具体作用机制还不清楚。简单的问题是什么是雌激素的作用，以及如何使雌激素在体内发挥作用。在本实验中，我们重点研究了重组腺病毒载体和/或条件转基因小鼠在体内获得的功能，这些重组腺病毒载体和/或条件转基因小鼠分别表达了人ERα和ERβ的构成活性，如CA ERα和ERβ。另一种策略是使用带有Cre/loxP系统的转基因小鼠来“有条件地获得功能”。这些转基因产物在wo/Cre重组酶处理前分别表达报告基因(nlsGFP/DsRed2)，处理后分别在Caerα和Caerβ中表达。通过重组Cre，可以选择性地激活时间和器官中的雌激素信号。最后，我们的目标是通过体内“选择性获得/丧失功能”来揭示雌激素信号的生理作用。在这个项目中，我们证明了EFP(雌激素反应性手指蛋白)是乳腺肿瘤细胞增殖所必需的，它通过蛋白分解14-3-3西格玛，这是细胞周期的负调节(自然，417,871-875,2002)。

项目成果

Fujita, M, Ogawa, S, Tsukui, T, et al., (他5名, 4番目): "Differential expression of secreted frizzled-related protein 4 in decidual cells during pregnancy"Journal of Mol. Endocrinol.. 28. 213-223 (2002)

Fujita, M, Okawa, S, Tsukui, T, et al.，（其他 5 人，第 4 期）：“怀孕期间蜕膜细胞中分泌的卷曲相关蛋白 4 的差异表达”杂志内分泌学.. 28. 213- 223（2002）

Tsukui, T., Toyoda, Y: "Methods in Molecuar Biology "Transgenic Techniques""Humana Press(In Press). (2002)

Tsukui，T.，Toyoda，Y：“分子生物学方法“转基因技术””Humana Press（正在出版）。

Fukuda, A, Tokonabe, S, Tsukui, T, et al., (他4名, 5番目): "Alleviation of PC4-mediated transcriptional repression by the ERCC3 helicase activity of general transcription factor TFHH"Journal of Biological Chemistry. (in press). (2003)

Fukuda, A, Tokonabe, S, Tsukui, T, et al.，（其他 4 人，第 5）：“一般转录因子 TFHH 的 ERCC3 解旋酶活性减轻 PC4 介导的转录抑制”，《生物化学杂志》（2003 年）。 ）

Urano, T, Saito, T, Tsukui, T, et al., (他5名, 3番目): "Efp targets 14-3-3sigma for proteolysis and promotes breast tumour growth"Nature. 417. 871-875 (2002)

Urano, T、Saito, T、Tsukui, T 等人，（其他 5 人，第 3 次）：“Efp 靶向 14-3-3sigma 进行蛋白水解并促进乳腺肿瘤生长”Nature 417. 871-875 (2002)。

Fujita, M, Urano, T, Tsukui, T, et al., (他6名, 5番目): "Estrogen activates cyclin-dependent kinases 4 and 6 through induction of cyclin D in rat primary osteoblasts"Biochem. Biophys. Res. Commun.. 299. 222-228 (2002)

Fujita, M, Urano, T, Tsukui, T, et al.，（其他 6 人，第 5）：“雌激素通过诱导大鼠原代成骨细胞中的细胞周期蛋白 D 激活细胞周期蛋白依赖性激酶 4 和 6”Biochem。 . 299. 222-228 (2002)