Early Growth Response-1 Regulates Angiopoietin-1-Induced Endothelial Cell Proliferation, Migration, and Differentiation
Early Growth Response-1 Regulates Angiopoietin-1-Induced Endothelial Cell Proliferation, Migration, and Differentiation
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DOI：
10.1161/atvbaha.108.181073
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发表时间：
2009-02-01
影响因子：
8.7
通讯作者：
Hussain, Sabah N. A.
中科院分区：
文献类型：
--
作者：
Abdel-Malak, Nelly A.;Mofarrahi, Mahroo;Hussain, Sabah N. A.
目的：血管生成素-1 （angiopoietin -1）是内皮细胞血管生成的重要调节因子。它促进细胞的迁移、增殖和分化，尽管参与这些过程的调节因子尚不清楚。在这项研究中，我们评估了转录因子早期生长反应-1 （Egr-1）对ang -1诱导的人脐静脉内皮细胞（HUVECs）血管生成的贡献。方法与结果：采用实时荧光定量PCR和免疫印迹法检测Egr-1的表达，采用电泳迁移位移法检测Egr-1的DNA结合活性。通过伤口愈合和Boyden室实验测量细胞迁移，而通过细胞计数、BrdU掺入和Matrigels来监测细胞增殖和细胞向毛细血管样管结构的分化。为了选择性地抑制Egr-1的表达，我们使用了siRNA寡核苷酸和特异性DNAzymes。Egr-1 mRNA的表达在暴露于Ang-1的2小时内上升了约9倍，此后下降。Egr-1表达的上调伴随着核动员的增加和DNA结合的增强。这些过程通过Erk1/2、PI-3激酶/AKT和mTOR途径介导。下调Egr-1表达完全消除了Ang-1诱导的内皮细胞迁移，显著降低了过表达Ang-1的HUVECs的增殖和毛细血管样管形成。结论- ang -1可触发Egr-1的瞬时显著诱导，Egr-1参与了ang -1诱导的内皮细胞迁移和增殖。（中华动脉血管杂志，2009;29:209-216）
Objective-Angiopoietin-1 (Ang-1) is an important regulator of angiogenesis in endothelial cells. It promotes migration, proliferation, and differentiation of cells, although the regulating factors involved in these processes remain unclear. In this study, we evaluated the contribution of the transcription factor early growth response-1 (Egr-1) to Ang-1-induced angiogenesis in human umbilical vein endothelial cells (HUVECs).Methods and Results-Expression of Egr-1 was evaluated with real-time PCR and immunoblotting, whereas Egr-1 DNA binding activity was monitored with electrophoretic mobility shift assays. Cell migration was measured with wound healing and Boyden chamber assays, whereas cell proliferation and differentiation of cells into capillary-like tube structures were monitored with cell counting, BrdU incorporation and Matrigels. To selectively inhibit Egr-1 expression, we used both siRNA oligonucleotides and specific DNAzymes. Egr-1 mRNA expression rose approximately 9-fold within 2 hours of Ang-1 exposure and declined thereafter. Upregulation of Egr-1 expression was accompanied by an increase in nuclear mobilization and augmented DNA binding. These processes were mediated through the Erk1/2, PI-3 kinase/AKT, and mTOR pathways. Knockdown of Egr-1 expression completely abrogated Ang-1-induced endothelial migration and significantly reduced proliferation and capillary-like tube formation of HUVECs that overexpress Ang-1.Conclusion-Ang-1 triggers significant and transient induction of Egr-1, and Egr-1 contributes to Ang-1-induced endothelial cell migration and proliferation. (Arterioscler Thromb Vasc Biol. 2009; 29: 209-216.)