Membrane binding sites and non-genomic effects of estrogen in cultured human pre-osteoclastic cells
Membrane binding sites and non-genomic effects of estrogen in cultured human pre-osteoclastic cells
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DOI：
10.1016/s0960-0760(96)00092-1
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发表时间：
1996-10-01
影响因子：
4.1
通讯作者：
Brandi, ML
中科院分区：
文献类型：
--
作者：
Fiorelli, G;Gori, F;Brandi, ML
除了功能性雌激素受体外，17 β-雌二醇的信号细胞表面结合位点的存在据报道，17 β E（2）在成骨细胞和破骨细胞样细胞中存在，表明17 β E（2）可能通过双重作用机制影响骨重塑：为了影响由类固醇-受体复合物的核活性介导的基因表达，并启动由细胞表面上的信号产生受体触发的快速反应。最近，我们证明了人前骨细胞系FLG 29.1具有功能性雌激素受体。在这项研究中，我们检查了FLG 29.1细胞表面是否存在17 β E（2）的结合位点，以及17 β E（2）是否可以引发细胞信号传导。使用细胞不渗透和荧光雌激素结合物，17 β-雌二醇-6-羧甲基肟-牛血清白蛋白-异硫氰酸荧光素，我们证明了17 β E特异性质膜结合位点的存在（2）。用低剂量（1 nM）和高剂量（1 μ M）的17 β E（2）刺激FLG 29.1细胞，诱导细胞pH迅速显著（P < 0.05）升高，这是使用图像分析系统在单细胞中测量的。此外，cAMP和cGMP均显著增加17 β E（2），并呈剂量依赖性反应。最后，使用图像分析系统在单细胞中测量，1 nM 17 β E（2）也诱导细胞内钙离子浓度[Ca 2 +]快速增加。我们的研究结果强烈提示17 β E（2）对破骨细胞前体的非基因组作用。版权所有（C）1996 Elsevier Science Ltd.
Besides functional estrogen receptors, the presence of signalling cell surface binding sites for 17 beta-estradiol (17 beta E(2)) has been reported in osteoblast- and osteoclast-like cells, suggesting that 17 beta E(2) may influence bone remodelling by a dual mechanism of action: to affect gene expression mediated by the nuclear activity of the steroid-receptor complex, and to initiate rapid responses triggered by a signal-generating receptor on the cell surface. Recently, we demonstrated that the human preosteoclastic cell line FLG 29.1 bears functional estrogen receptors. In this study we examined FLG 29.1 cells for the presence of cell surface binding sites for 17 beta E(2), and whether 17 beta E(2) could elicit cell signalling. Using a cell-impermeant and fluorescent estrogen conjugate, 17 beta-estradiol-6-carboxymethyloxime-bovine serum albumin-fluorescein isothiocyanate, we demonstrated the presence of specific plasma membrane binding sites for 17 beta E(2). Stimulation of FLG 29.1 cells with low (1 nM) and high (1 mu M) doses of 17 beta E(2) induced a prompt and significant (P < 0.05) increase of cellular pH, as measured in single cells using an image analysis system. In addition, both cAMP and cGMP were significantly increased by 17 beta E(2) with a dose-dependent response. Finally, a rapid increase of intracellular calcium ion concentration [Ca2+] was also induced by 1nM 17 beta E(2), as measured in single cells using an image analysis system. Our findings strongly suggest a non-genomic action of 17 beta E(2) on osteoclast precursors. Copyright (C) 1996 Elsevier Science Ltd.