Glucocorticoid regulation of human eosinophil gene expression
Glucocorticoid regulation of human eosinophil gene expression
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DOI：
10.1016/s0960-0760(03)00065-7
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发表时间：
2003-03-01
影响因子：
4.1
通讯作者：
Miesfeld, RL
中科院分区：
文献类型：
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作者：
Chauhan, S;Leach, CH;Miesfeld, RL
在新鲜分离的人嗜酸性粒细胞的类固醇调节的基因表达的分子分析是困难的，由于固有的自发凋亡率高，内源性核糖核酸酶的水平升高。为了规避这些限制，我们确定了人嗜酸性细胞系EoL-1是否可以作为糖皮质激素信号转导的体外模型。通过优化低血清培养基中的生长条件，我们发现地塞米松（Dex）处理EoL-1细胞诱导的凋亡途径被白细胞介素-5（IL-5）抑制。此外，使用未经处理的EoL-1细胞和新鲜分离的人嗜酸性粒细胞的RNA进行基因表达谱分析，确定了380个常见表达的基因，包括嗜酸性粒细胞标志物颗粒主要碱性蛋白，异黄酮内过氧化物合酶I和花生四烯酸15-脂氧合酶。使用地塞米松处理0、4、12、24和48 h的EoL-1细胞进行表达谱分析，鉴定出162个差异表达的基因。基于表达谱的两个最高度上调的基因是转录因子Ets-2和MHC 11类基因（Q、R和P）。EoL-1细胞中这些基因的表达在RNA和蛋白质水平上被地塞米松诱导，这与Ets-2在控制细胞周期进程中的已知功能以及MHC II类抗原在介导嗜酸性粒细胞功能中的作用一致。(C)2003爱思唯尔科技有限公司版权所有。
Molecular analysis of steroid-regulated gene expression in freshly isolated human eosinophils is difficult due to the inherent high rate of spontaneous apoptosis and elevated levels of endogenous ribonucleases. To circumvent these limitations, we determined if the human eosinophilic cell line EoL-1 could serve as an in vitro model of glucocorticoid signaling. We found by optimizing growth conditions in low serum-containing, media that dexamethasone (Dex) treatment of EoL-1 cells induced an apoptotic pathway that was inhibited by interleukin-5 (IL-5). Moreover, gene expression profiling using RNA from untreated EoL-1 cells and from freshly isolated human eosinophils identified 380 commonly expressed genes, including the eosinophil markers granule major basic protein, prostaglandin-endoperoxide synthase I and arachidonate 15-lipoxygenase. Expression profiling was performed using EoL-1 cells that had been treated with dexamethasone for 0, 4, 12, 24 and 48 h identifying 162 genes as differentially expressed. Two of the most highly upregulated genes based on expression profiling were the transcription factor Ets-2 and the MHC Class 11 genes (Q, R, and P). Expression of these genes in EoL-1 cells was shown to be dexamethasone-induced at the RNA and protein levels which is consistent with the known function of Ets-2 in controlling cell cycle progression and the role of MHC Class II antigens in mediating eosinophil functions. (C) 2003 Elsevier Science Ltd. All rights reserved.