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Non-genomic actions of estrogen in breast cancer

雌激素在乳腺癌中的非基因组作用

基本信息

批准号：
7407543
负责人：
ELLIS R LEVIN
金额：
$ 21.76万
依托单位：
SOUTHERN CALIFORNIA INST FOR RES/EDUC
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-08-02 至 2009-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7407543
关键词：
1-Phosphatidylinositol 3-Kinase Aliquot BRCA1 Protein BRCA1 gene Binding Biological Breast Cancer Cell Cell Cycle Cell Cycle Progression Cell Cycle Stage Cell Nucleus Cell Proliferation Cell Survival Cell membrane Cells Condition Cultured Cells Cyclin D1 Cyclins DTR gene DUSP1 gene Dimerization Dominant-Negative Mutation EGF gene Epidermal Growth Factor Receptor Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor Estradiol Estrogen Nuclear Receptor Estrogen Receptors Estrogens Event Excision Family Fatty acid glycerol esters GTP Binding GTP-Binding Protein alpha Subunits GTP-Binding Proteins Genetic Transcription Genomics Goals Growth Hereditary Breast Carcinoma Human Immunoprecipitation In Situ In Vitro Insulin-Like Growth Factor I Lead Link Location M cell MCF7 cell MEKs Malignant Neoplasms Mammary gland Matrix Metalloproteinases Measures Mediating Membrane Mitogen-Activated Protein Kinases Modeling Nuclear Nuclear Envelope Nuclear Receptors Nude Mice PD-98059 Phosphoinositide-3-Kinase, Catalytic, Gamma Polypeptide Phosphoric Monoester Hydrolases Phosphotransferases Production Protein Overexpression Proteins Publishing Radiolabeled Receptor Cell Receptor Cross-Talk Receptor Signaling Risk Signal Transduction Small Interfering RNA Stimulation of Cell Proliferation System Testing Transactivation Tyrphostin AG 825 Up-Regulation Western Blotting angiogenesis base cell type day dimer diphtheria toxin receptor heparin-binding EGF-like growth factor in vivo inhibitor/antagonist malignant breast neoplasm member migration mutant non-genomic novel prevent radiotracer receptor receptor function response tumor tumor growth wortmannin yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Estrogen enhances the risk of developing breast cancer. Non-genomic effects of estrogen result from signal transduction originating at the membrane, in part from cross talk between ER and the EGF receptor. Cell based systems will establish that blocking E2-signaling through ERK and Pl3 kinase for 3 days substantially prevents the proliferation of cultured MCF-7 and ZR-75-1 (ER+) cells. G1 cell cycle targets and GI/S progression are ERK and Pl3K entrained. It is proposed that E2 interacts with BRCA1 to promote breast cancer. The ability of E2 to activate ERK will be shown to be down regulated by expression of wild type but not mutant BRCA1 in HCC-1937 cells (express a mutant BRCA1), co-transfected to express ER. This model will also determine whether wtBRCA1 can block the proliferative and ERK-inducing effects of E2, in cells cultured for 3 days. We will implicate the membrane receptor by expressing a dominant negative ER that only abrogates endogenous membrane ER function, and by targeting the E domain to the membrane. It is proposed that E2 binds the membrane ER, activates discrete G proteins that activate Src. Src activates specific matrix metalloproteinases that liberate HB-EGF and transactivates the EGFR or ErbB2. This leads to ERK and PI3K activation in breast cancer cells. This will be shown using dominant negative constructs SiRNA, and cells deficient for EGFR or ErbB2. We propose that the receptor must also dimerize to signal from the membrane, and we will use expression of a dimer mutant to show this. In-vivo, E domain targeted to membrane or nucleus are stably expressed in ER - breast cancer cells. These cells are injected into nude mice, and growth under estrogen-repleted conditions is determined. The goal is to define non-genomic actions of estrogen that could potentially be antagonized at the membrane, yet preserve the desirable effects of E2 in the nucleus.

描述（由申请人提供）：雌激素增加患乳腺癌的风险。雌激素的非基因组效应是由起源于膜的信号传导引起的，部分原因是ER和EGF受体之间的串扰。基于细胞的系统将确定阻断通过ERK和P13激酶的E2-信号传导3天基本上防止了培养的MCF-7和ZR-75-1（ER+）细胞的增殖。G1细胞周期靶点和GI/S进展是ERK和P13 K夹带的。研究表明，E2与BRCA 1相互作用促进乳腺癌的发生。在共转染表达ER的HCC-1937细胞（表达突变BRCA 1）中，E2激活ERK的能力将被野生型而非突变BRCA 1的表达下调。该模型还将确定在培养3天的细胞中，wtBRCA 1是否可以阻断E2的增殖和ERK诱导作用。我们将通过表达一个显性负性ER，仅消除内源性膜ER功能，并通过靶向E结构域的膜牵连的膜受体。有人提出，E2结合膜ER，激活离散的G蛋白，激活Src。Src激活释放HB-EGF的特异性基质金属蛋白酶，并反式激活EGFR或ErbB 2。这导致乳腺癌细胞中的ERK和PI 3 K活化。这将使用显性阴性构建体SiRNA和EGFR或ErbB 2缺陷的细胞来显示。我们提出，受体也必须二聚化，从膜的信号，我们将使用二聚体突变体的表达来证明这一点。在体内，靶向膜或核的E结构域在ER -乳腺癌细胞中稳定表达。将这些细胞注射到裸鼠中，并测定在雌激素充足条件下的生长。目的是确定雌激素的非基因组作用，这些作用可能在膜上被拮抗，但在细胞核中保留了E2的理想作用。