权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Analysis of the mechanism of estrogen-dependent proliferation of endometrial carcinoma cells via IGF-1/MAPK

IGF-1/MAPK分析子宫内膜癌细胞雌激素依赖性增殖机制

基本信息

批准号：
18591830
负责人：
ASHIDA Takashi
金额：
$ 2.57万
依托单位：
Shinshu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591830/
关键词：
endometrial carcinoma estrogen MAP kinasae IGF-1 cychn MAP Kinase

项目摘要

To examine estrogen-induced growth mechanisms of endometrial carcinoma, we investigated the estrogen-induced activation of the mitogen-activated protein kinase (MAPK) pathway and cell cycle regulators. Estradiol (E2) treatment at concentrations of 10^8M and 10^6M to estrogen receptor (ER)-positive endometrial carcinoma Ishikawa cells for 24 hours resulted in increased cell proliferation by 20% and 28%, respectively. The E2-induced proliferation was associated with activation of extracellular signal-regulated kinase (ERK)1/2 and up-regulation of cyclin Dl and E, which were suppressed by the addition of a MEK inhibitor (U0126) or an ER antagonist (ICI182,780). Then, our screening for estrogen-inducible growth factors identified that insulin-like growth factor (IGF)-1 was up-regulated remarkably by E2. Immunoprecipitation using conditioned medium of Ishikawa cells after E2 treatment confirmed the E2-induced secretion of IGF-1 protein. Treatment with recombinant IGF-1 stimulated cell proliferation in a dose-dependent fashion, in association with ERK1/2 phosphorylation and up-regulation of cyclin D1 and E. These IGF-1-induced responses were suppressed by treatment with ER antagonist, MEK inhibitor, or anti-IGF-1 receptor antibody. Immunohistochemical staining confirmed the expression of activated ERK1/2 in normal proliferative phase endometria and endometrial carcinomas, indicating the involvement of this pathway in actively proliferating endometrial tissues in vivo. These findings suggest that E2-induced proliferation of endometrial carcinoma cells is mediated by the ERK1/2 pathway via autocrine stimulation of IGF- 1.

为了探讨雌激素诱导子宫内膜癌生长的机制，我们研究了雌激素诱导的丝裂原活化蛋白激酶(MAPK)通路和细胞周期调节因子的激活。雌二醇(E_2)作用于雌激素受体(ER)阳性的子宫内膜癌Ishikawa细胞24小时，细胞增殖分别增加20%和28%。ER拮抗剂(ICI182,780)或MEK抑制剂(U0126)可抑制细胞外信号调节激酶(ERK)1/2的激活，上调细胞周期蛋白D1和E的表达。然后，我们对雌激素诱导的生长因子的筛选发现，胰岛素样生长因子(IGF)-1被E2显著上调。经E2处理后的Ishikawa细胞条件培养液的免疫沉淀证实了E2诱导的IGF-1蛋白的分泌。重组IGF-1以剂量依赖的方式刺激细胞增殖，并伴随着ERK1/2的磷酸化和细胞周期蛋白D1和E的上调，这些反应可被ER拮抗剂、MEK抑制剂或抗IGF-1受体抗体抑制。免疫组织化学染色证实激活的ERK1/2在正常增生期子宫内膜和子宫内膜癌中均有表达，提示该通路参与了活体子宫内膜组织的活跃增殖。这些发现表明，雌激素诱导的子宫内膜癌细胞的增殖是通过ERK1/2途径通过IGF-1的自分泌刺激而介导的。