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【ATVB】早期生长反应蛋白1调节了血管生成素1诱
(Arteriosclerosis, Thrombosis, and Vascular Biology. 2009;29:202.)
Early Growth Response-1 Regulates Angiopoietin-1-Induced Endothelial Cell Proliferation, Migration, and Differentiation
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 bindingactivity 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 nuclearmobilization 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. Early Growth Response-1 Regulates Angiopoietin-1-Induced Endothelial Cell Proliferation, Migration, and Differentiation
早期生长反应蛋白1调节了血管生成素1诱导的内皮细胞的增殖、迁移和分化
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).
目的：血管生成素1（Ang-1）是内皮细胞中重要的血管生成调节因子之一。它促进了细胞的迁移、增殖和分化，尽管该过程中牵涉的因子尚不清楚。在本研究中，我们评价了转录因子早期生长反应蛋白1（Egr-1）对Ang-1诱导的人类脐静脉内皮细胞（HUVECs）的血管生成的作用。
Methods and Results— Expression of Egr-1 was evaluated with real-time PCR and immunoblotting, whereas Egr-1 DNA bindingactivity 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 nuclearmobilization 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.
方法和结果：使用实时PCR和免疫印迹法对Egr-1的表达进行了评价，使用凝胶迁移滞后实验对Egr-1的DNA结合活性进行了监测。使用伤口愈合和Boyden小室检测法对细胞的迁移进行了测量，使用细胞计数、BrdU掺入和麦氏胶对细胞的增殖和细胞向毛细血管样小管结构的分化进行了监测。我们使用siRNA和特异性的DNA酶抑制了Egr-1的表达。Egr-1mRNA的表达，在细胞暴露于Ang-1中2小时后会增高大约9倍，随后下降。Egr-1表达的上升伴随着核动员[标签:content1][标签:content2]
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作者:admin@医学,生命科学 2011-03-05 05:11
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