Using U0126 to dissect the role of the extracellular signal-regulated kinase 1/2 (ERK1/2) cascade in the regulation of gene expression by endothelin-1 in cardiac myocytes
Using U0126 to dissect the role of the extracellular signal-regulated kinase 1/2 (ERK1/2) cascade in the regulation of gene expression by endothelin-1 in cardiac myocytes
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DOI：
10.1016/j.yjmcc.2006.04.011
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发表时间：
2006-08-01
影响因子：
5
通讯作者：
Clerk, Angela
中科院分区：
文献类型：
--
作者：
Kennedy, Robert A.;Kemp, Timothy J.;Clerk, Angela
肥厚激动剂内皮素-1 快速但短暂地激活心肌细胞中的细胞外信号调节激酶 1/2 (ERK1/2) 级联（和其他信号通路），但将其与肥厚联系起来的事件尚不清楚。使用 Affymetrix 大鼠 U34A 微阵列，我们鉴定了内皮素-1 单独或与 ERK1/2 级联抑制剂 U0126 联合诱导的新生儿肌细胞基因表达的短期（2-4 小时）变化。单独使用 U0126 显着改变了 15 个基因的表达，内皮素 1 显着改变了另外 78 个基因的表达。在 U0126 上调的基因中，有 4 个通常是由二恶英通过芳基碳氢化合物受体 (AhR) 诱导的，这表明 U0126 激活心肌细胞中的异生物质反应元件，可能与对 ERKI/2 信号传导的影响无关。显示内皮素-1 表达改变的 78 个基因形成了 5 个簇：(i) 三个簇显示内皮素-1 根据时间进程（4 小时 > 2 小时；2 小时 > 4 小时；2 小时与 4 小时相似）上调，且至少被 U0126 部分抑制； (ii) 一组 11 个基因被内皮素-1 上调，但不受 U0126 影响，表明通过 ERK1/2 以外的信号通路进行调节； (iii) 一组六个基因被内皮素-1 下调，并被 U0126 减弱。因此，UO 126 显然激活了心肌细胞中的 AhR（在长期研究中必须考虑到这一点），但仔细的分析可以识别出可能通过 ERK1/2 级联急剧调节的基因。我们的数据表明，内皮素-1 诱导的基因表达的大部分变化是由 ERKI/2 级联介导的。 (c) 2006 Elsevier Inc. 保留所有权利。
The hypertrophic agonist endothelin-1 rapidly but transiently activates the extracellular signal-regulated kinase 1/2 (ERK1/2) cascade (and other signalling pathways) in cardiac myocytes, but the events linking this to hypertrophy are not understood. Using Affymetrix rat U34A microarrays, we identified the short-term (2-4 h) changes in gene expression induced in neonatal myocytes by endothelin-1 alone or in combination with the ERK1/2 cascade inhibitor, U0126. Expression of 15 genes was significantly changed by U0126 alone, and expression of an additional 78 genes was significantly changed by endothelin-1. Of the genes upregulated by U0126, four are classically induced through the aryl hydrocarbon receptor (AhR) by dioxins suggesting that U0126 activates the xenobiotic response element in cardiac myocytes potentially independently of effects on ERKI/2 signalling. The 78 genes showing altered expression with endothelin-1 formed five clusters: (i) three clusters showing upregulation by endothelin-1 according to time course (4 h > 2 h; 2 h > 4 h; 2 h similar to 4 h) with at least partial inhibition by U0126; (ii) a cluster of 11 genes upregulated by endothelin-1 but unaffected by U0126 suggesting regulation through signalling pathways other than ERK1/2; (iii) a cluster of six genes downregulated by endothelin-1 with attenuation by U0126. Thus, UO 126 apparently activates the AhR in cardiac myocytes (which must be taken into account in protracted studies), but careful analysis allows identification of genes potentially regulated acutely via the ERK1/2 cascade. Our data suggest that the majority of changes in gene expression induced by endothelin-1 are mediated by the ERKI/2 cascade. (c) 2006 Elsevier Inc. All rights reserved.