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首页> 外文期刊>Journal of Cellular Physiology >A high throughput method to study the physiology of E2:ERα signaling in breast cancer cells
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A high throughput method to study the physiology of E2:ERα signaling in breast cancer cells

机译：高通量方法研究生理学E2:在乳腺癌细胞ERα信号

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17β‐estradiol (E2) regulates diverse physiological effects including cell proliferation through the estrogen receptor α (ERα), which as a transcription factor drives gene transcription and as an extra‐nuclear localized receptor triggers the membrane‐dependent activation of diverse kinase cascades. E2 also modifies ERα intracellular levels via diverse intracellular mechanisms. In this way, the E2‐acivated ERα integrates signaling cascades with the modulation of receptor intracellular concentration and with the induction of DNA synthesis and ultimately drives cell proliferation. In turn, E2 signaling deregulation can cause many diseases including breast cancer (BC). Recently, we performed a Western blotting (WB)‐based screen to identify novel pathways affecting ERα intracellular levels and BC cell proliferation. However, because WB lacks high throughput potential, a high‐content method to detect all aspects of E2:ERα signaling (nuclear and extra‐nuclear receptor activity, ERα levels, E2‐induced DNA synthesis) is desirable. Here, we set up a rapid way to measure E2:ERα signaling in 96‐well plate format. To demonstrate its robustness, we also challenged 4OH‐tamoxifen resistant (Tam‐Res) BC cells with a library of anti‐cancer drugs and identified methotrexate (MTX) as a molecule inducing ERα degradation and preventing BC cell proliferation. Overall, our research provides a high‐content technique to study the physiology of E2:ERα signaling in cells and further suggests a possible anti‐ERα and anti‐proliferative use for MTX in Tam‐Res BCs.
机译：17β雌二醇(E2)调节各种生理包括细胞增殖的影响雌激素受体α(ERα),这是一个转录因子基因转录作为一个额外的核本地化的受体触发膜相关的激活不同的激酶级联。通过不同的胞内细胞内水平机制。集成与调制信号级联受体细胞内浓度和DNA合成,最终的感应细胞增殖。放松管制会导致许多疾病,包括乳腺癌(BC)。西方墨点法(WB)基于列车屏幕来定义新颖的途径影响ERα细胞内水平公元前和细胞增殖。缺乏高吞吐量的潜力,一个高的内容方法检测E2的所有方面:ERα信号(核和额外的核受体活性,ERα水平,E2诱导DNA合成)是可取的。在这里,我们建立了一个快速的方法来测量E2: ERα96年信号量板格式。其鲁棒性,我们也挑战4哦它莫西芬耐药(公元前Tam Res)应承担的细胞库的抗癌症药物和确定甲氨蝶呤(MTX)作为分子诱导ERα退化和防止BC细胞增殖。研究提供了一个高技术的内容研究E2的生理学:ERα信号在细胞并进一步提出一个可能的反ERα和抗增殖使用MTX在Tam中应承担的Res bc。

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