DOMINANT NEGATIVE ESTROGEN RECEPTORS AND BREAST CANCER

显性负雌激素受体与乳腺癌

• 批准号: 2112662
• 负责人: KIRK E EKENA
• 金额: $ 2.26万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-12-20 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2112662
• 关键词: DOMINANT; NEGATIVE; ESTROGEN; RECEPTORS; BREAST

The objective of this research is to develop and investigate a novel method for functional inactivation of estrogen receptor (ER) in estrogen-responsive human breast cancer cells. The approach is based on dominant negative ER mutants which, while inactive on their own, suppress the activity of wildtype ER when they are co-expressed in the same cells. Powerful ER mutants will be generated by replacing the N- and C-terminal transactivation domains with polyalanine tracts and by using the P22 challenge phage system to isolate mutations that result in increased ability of mutant ER to compete for binding to estrogen response element DNA. The relative contributions of competition for binding to the ERE, formation of heterodimers, and interference with ER- specific transcription components to the activity of the dominant negative mutants will be determined. Stably transfected MCF-7 human breast cancer cells producing the dominant negative mutant ERs will be examined for suppression of estrogen-stimulated growth in vitro, and in vivo using athymic nude mice. These studies will provide new insights into ER regulation of gene expression and the proliferation and tumorigenicity of breast cancer cells, and allow evaluation of a novel approach for the extinction of estrogen-dependent breast cancer growth.

本研究的目的是开发和研究一种新的 雌激素受体（ER）功能性失活的方法 雌激素敏感的人乳腺癌细胞。该方法是基于 显性负性ER突变体，虽然本身无活性， 抑制野生型ER的活性， 同样的细胞。强大的ER突变体将通过取代N- 和C-末端反式激活结构域与多聚丙氨酸区， 使用P22攻击噬菌体系统来分离导致 增强突变ER竞争结合雌激素的能力 响应元件DNA竞争的相对贡献 与ERE结合，形成异源二聚体，并干扰ER- 特异性转录成分对主导活性的影响 将确定阴性突变体。稳定转染的MCF-7人 产生显性负突变ER的乳腺癌细胞， 检查体外雌激素刺激生长的抑制作用， 体内使用无胸腺裸鼠。这些研究将提供新的见解 进入ER调节基因表达和增殖， 乳腺癌细胞的致瘤性，并允许评估一种新的 雌激素依赖性乳腺癌生长的灭绝方法。