Expression and regulation of the estrogen receptors in PC-3 human prostate cancer cells
Expression and regulation of the estrogen receptors in PC-3 human prostate cancer cells
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DOI：
10.1016/j.steroids.2015.12.021
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发表时间：
2016-03-01
期刊：
影响因子：
2.7
通讯作者：
Porto, C. S.
中科院分区：
文献类型：
--
作者：
Pisolato, R.;Lombardi, A. P. G.;Porto, C. S.
本研究的目的是确定在雄激素非依赖性前列腺癌细胞系PC-3中经典雌激素受体ER α、ER β、ER-α 36亚型和GPER的表达、细胞定位和调节。此外，我们评估了这些受体对ERK 1/2（细胞外信号调节蛋白激酶）信号通路的激活的相对贡献。这四种雌激素受体通过Western印迹法检测，并通过免疫荧光法显示优先定位于PC-3细胞的胞核区域。此外，用17 β-雌二醇（E2）（1 μ M）处理24小时导致经典雌激素受体的下调，而E2在生理浓度（0.1 nM）处理24小时往往会增加ER α和ER β的水平。此外，ER α选择性激动剂PPT选择性增加ER β的表达，ER β选择性激动剂DPN增加ER α水平。这些处理均不影响ER-α 36亚型的表达。这些细胞中经典雌激素受体的不寻常的细胞质定位不同于大多数雌激素靶细胞中的核定位，并表明快速信号传导途径可能被优先激活。事实上，用ER α、ER β和GPER的选择性激动剂处理诱导ERK 1/2磷酸化，其被各自的拮抗剂阻断。另一方面，E2诱导的ERK 1/2激活可能涉及其他机制，因为它不被三种拮抗剂阻断。综上所述，这些结果表明ER α和ER β之间存在相互作用，以调节彼此的表达，并表明其他受体，如ER-α 36，参与了E2对ERK 1/2的快速激活。识别新的ER亚型，调节受体和信号通路对于开发去势抵抗性前列腺癌的新治疗策略具有重要意义。(C)2016 Elsevier Inc. All rights reserved.
The aim of this study was to identify the expression, cellular localization and regulation of classic estrogen receptors ER alpha, and ER beta, ER-alpha 36 isoform and GPER in the androgen-independent prostate cancer cell line PC-3. In addition, we evaluated the relative contribution of these receptors to the activation of the ERK1/2 (extracellular signal-regulated protein kinases) signaling pathway.These four estrogen receptors were detected by Western blot assays and were shown by immunofluorescence assays to localize preferentially in extranuclear regions of PC-3 cells. In addition, treatment with 17 beta-estradiol (E2) (1 mu M) for 24 h led to down-regulation of the classic estrogen receptors, whereas E2 at physiological concentration (0.1 nM) for 24 h tended to increase the levels of ER alpha and ER beta. Furthermore, the ER alpha-selective agonist PPT selectively increased the expression of ER beta and the ER beta-selective agonist DPN increased ER alpha levels. None of these treatments affected expression of the ER-alpha 36 isoform. The unusual cytoplasmic localization of the classic estrogen receptors in these cells differs from the nuclear localization in the majority of estrogen target cells and suggests that rapid signaling pathways may be preferentially activated. In fact, treatment with selective agonists of ER alpha, ER beta and GPER induced ERK1/2 phosphorylation that was blocked by the respective antagonists. On the other hand, activation of ERK1/2 induced by E2 may involve additional mechanisms because it was not blocked by the three antagonists. Taken together, the results indicate that there is a crosstalk between ER alpha and ER beta to regulate the expression of each other, and suggest the involvement of other receptors, such as ER-alpha 36, inthe rapid ERK1/2 activation by E2. The identification of new isoforms of-ERs, regulation of the receptors and signaling pathways is important to develop new therapeutic strategies for the castration-resistant prostate cancer. (C) 2016 Elsevier Inc. All rights reserved.