Tikkanen M K, Carter D J, Harris A M, Le H M, Azorsa D O, Meltzer P S, Murdoch F E
The Hospital of Loimaa, 32201 Loimaa, Finland.
Proc Natl Acad Sci U S A. 2000 Nov 7;97(23):12536-40. doi: 10.1073/pnas.220427297.
Coactivators are believed to mediate estrogen-induced gene responses via interaction with estrogen receptors (ER). Currently, a major challenge is to determine the importance of each coactivator in a specific cell type and promoter context in response to a particular ligand. The potential of ER to interact with a growing list of coactivators has been shown in a variety of in vitro and gene transfer assays, yet very few data have demonstrated the interaction of endogenous coactivators with ER in intact cells. We report here a ligand-specific interaction of endogenous human ER (hER) and the AIB1 coactivator in MCF-7 human breast cancer cells by using immunoprecipitation analyses. Complexes between endogenously expressed hER and AIB1 were detected in estradiol-treated cells and to a much lesser extent in cells treated with the partial agonist, monohydroxytamoxifen. We were unable to detect an hER-SRC-1 complex in our immunoprecipitations from MCF-7 cells. The in vitro-binding affinity for mouse ER interaction with AIB1 was estimated to be 40-120 nM. We conclude that AIB1 is a major coactivator for hER in MCF-7 human breast cancer cells.
人们认为共激活因子通过与雌激素受体(ER)相互作用来介导雌激素诱导的基因反应。目前,一个主要挑战是确定在特定细胞类型和启动子背景下,每种共激活因子在响应特定配体时的重要性。在各种体外和基因转移试验中,已显示出ER与越来越多的共激活因子相互作用的潜力,但很少有数据证明内源性共激活因子与完整细胞中的ER相互作用。我们在此报告,通过免疫沉淀分析,在MCF-7人乳腺癌细胞中内源性人ER(hER)与AIB1共激活因子存在配体特异性相互作用。在雌二醇处理的细胞中检测到内源性表达的hER与AIB1之间的复合物,而在用部分激动剂单羟基他莫昔芬处理的细胞中检测到的复合物要少得多。在我们从MCF-7细胞进行的免疫沉淀中,未能检测到hER-SRC-1复合物。据估计,小鼠ER与AIB1的体外结合亲和力为40-120 nM。我们得出结论,AIB1是MCF-7人乳腺癌细胞中hER的主要共激活因子。