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首页> 外文期刊>Journal of cellular physiology. >Real-time measurement of E2: ERa transcriptional activity in living cells
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Real-time measurement of E2: ERa transcriptional activity in living cells

机译：实时测量E2:转录时代在活细胞的活动

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摘要

Kinetic analyses of diverse physiological processes have the potential to unveil new aspects of the molecular regulation of cell biology at temporal levels. 17β-estradiol (E2) regulates diverse physiological effects by binding to the estrogen receptor a (ERa), which primarily works as a transcription factor. Although many molecular details of the modulation of ERa transcriptional activity have been discovered including the impact of receptor plasma membrane localization and its relative E2-evoked signaling, the knowledge of real-time ERa transcriptional dynamics in living cells is lacking. Here, we report the generation of MCF-7 and HeLa cells stably expressing a modified luciferase under the control of an E2-sensitive promoter, which activity can be continuously monitored in living cells and show that E2 induces a linear increase in ERa transcriptional activity. Ligand-independent (e.g., epidermal growth factor) receptor activation was also detected in a time-dependent manner. Kinetic profiles of ERa transcriptional activity measured in the presence of both receptor antagonists and inhibitors of ERa plasma membrane localization reveal a biphasic dynamic of receptor behavior underlying novel aspects of receptor-regulated transcriptional effects. Finally, analysis of the rate of the dose-dependent E2 induction of ERa transcriptional activity demonstrates that low doses of E2 induce an effect identical to that determined by high concentrations of E2 as a function of the duration of hormone administration. Overall, we present the characterization of sensitive stable cell lines were to study the kinetic of E2 transcriptional signaling and to identify new aspects of ERa function in different physiological or pathophysiological conditions.
机译：不同的生理的动力学分析有可能推出新的过程方面的分子调控的细胞生物在时间的水平。调节各种生理效应绑定到雌激素受体(时代)主要作为转录因子。尽管许多分子调制的细节时代的转录活动发现包括受体的影响质膜定位及其相对的E2-evoked信号,实时的知识时代在活细胞转录动力学缺乏。和海拉细胞稳定表达修改荧光素酶E2-sensitive的控制启动子,可以不断地活动监控活细胞和表明E2诱发转录线性增加的时代活动。生长因子受体激活也发现时间的方式。的转录活动测量时代在这两种受体拮抗剂抑制剂的质膜定位时代揭示了受体的两相的动态行为receptor-regulated底层小说的方面转录的影响。时代的E2感应存在剂量依赖的相关性转录活动表明,低剂量的E2诱导效果相同由高浓度的E2作为决定的激素的持续时间的函数管理。特征敏感的稳定的细胞系研究E2转录的动能信号和识别方面的新纪元在不同的生理或函数病理生理条件。

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