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

Analysis of the molecular mechanisms of the estrogen-dependent growth of the endometrial carcinoma

子宫内膜癌雌激素依赖性生长的分子机制分析

基本信息

批准号：
12671587
负责人：
SHIOZAWA Tanri
金额：
$ 0.96万
依托单位：
Shinshu University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671587/
关键词：
endometrium endometrial carcinoma estrogen cyclin c-Jun cdk transcription factor C-Jun

项目摘要

The growth of the normal human endometrium and estrogen receptor ( ER ) -positive endometrial carcinoma is stimulated by estrogen via ER.Recent studied have revealed that the cell cycle-related molecules, such a cyclins, cyclin dependent kinases ( cdk ), and cdk inhabitors ( cdkI ) play essential roles in cell proliferation. The present study was undertaken to incestigate the possible involvement of the cell cycle-related molecules in estogen-dependent growth of the normal and neoplastic endometrium.Immunohistochemical staining demonstrated that in the normal endometrial glands, cyclins ( D1, E, A, B1 ) and cdks ( cdk4, cdk2, cdc2 ) were sporadically expressed in the proliferative phase, and were overexpressed in 30-75 % of the endometrialcarcanoma examined. Overexpression of cylin A and cdk4 wa correlated with patients' survival.The molecular mechanisms for the estrogen-induced growth of the endometriaum was analyzed using cultured glandular cells from normal proliferative endometrium … More with or without estradiol ( E_2 ) stimulation. Expression of cyclin D1 was detected 4 hours after E_2 treatment, and followed by serial expressions of cyclin E, A, and B1. For the intracellur pathway of cyclin D1 up-regulation by estrogen, we hypothesized the involvement of transcription factor, c-Jun/c-Fos, since te cyclin D1 posess c-Jun/c-Fos binding ( AP-1 ) site but lacks the estrogen responsive element. The expression of c-Jun in cultured endometrial cell occurred 2 hours after E_2 stimulation., preceding the cyclin D1 expression. Luciferase assay using various cyclin D1 promoter constructs revealed that the elecated luciferase activity afterE, stimulation was observed exclusively in the costructs possessing AP-1 site. Gel shift mobility assay indicated the binding of AP-1 bining site to nuclear extracts from E_2 treated endometrial epithelia. Increased luciferas activity was also observed when c-Jun expression plasmid was co-transfected with containing AP-1 site. On the other hand, inER-positive endometrial carcinoma ( Ishikawa ) cells, the expression pattern cyclins after E_2 stimulation as well as promoter activity of cyclin D1 differed from those of te normal endometrial cells. These findings seggested that the estrogen-stimulated expression of cyclin D1 in the normal endometrial glands is mediated by AP-1, and that was different from ER-positive endometrial carcinoma. Less

雌激素通过雌激素刺激正常子宫内膜和雌激素受体(ER)阳性的子宫内膜癌的生长。最近的研究表明，细胞周期相关分子，如细胞周期相关分子，如细胞周期蛋白、细胞周期蛋白依赖性激酶(CDK)和CDK抑制因子(CDKI)在细胞增殖中起重要作用。本研究旨在探讨细胞周期相关分子在正常和肿瘤子宫内膜雌激素依赖性生长中的作用。免疫组织化学染色显示，在正常子宫内膜腺组织中，细胞周期蛋白(d1、E、A、B1)和CDKs(cdk4、cdk2、cdc2)在增生期呈零星表达，在30-75%的子宫内膜癌中呈过表达。Cylin A和CDK4的过度表达与患者的生存有关。利用培养的正常增生期子宫内膜腺细胞…分析了雌激素诱导子宫内膜生长的分子机制有或无雌二醇(E_2)刺激的更多。E_2处理4h后检测细胞周期蛋白D1的表达，随后检测细胞周期蛋白E、A、B1的表达。对于雌激素上调细胞周期蛋白D1的细胞内途径，我们假设转录因子c-jun/c-Fos参与其中，因为细胞周期蛋白D1具有c-jun/c-Fos结合(AP-1)位点，但缺乏雌激素反应元件。C-jun在子宫内膜细胞中的表达出现在E_2刺激后2小时，先于细胞周期蛋白D_1表达。用不同的Cyclin D1启动子构建的荧光素酶分析表明，刺激后的荧光素酶活性仅在含有AP-1位点的结构中观察到。凝胶迁移率分析显示，E_2处理的子宫内膜上皮细胞核提取物与AP-1结合位点结合。将含有AP-1位点的c-jun表达载体与c-jun表达载体共转染后，荧光素酶活性也明显增强。另一方面，子宫内膜癌(Ishikawa)细胞的INER阳性表达、E_2刺激后细胞周期蛋白的表达模式以及细胞周期蛋白D1的启动子活性与正常子宫内膜细胞不同。这些结果表明，雌激素刺激的正常子宫内膜腺中细胞周期蛋白D1的表达是由AP-1介导的，这与ER阳性的子宫内膜癌不同。较少