Green fluorescent protein selectively induces HSP70-mediated up-regulation of COX-2 expression in endothelial cells
Green fluorescent protein selectively induces HSP70-mediated up-regulation of COX-2 expression in endothelial cells
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DOI：
10.1182/blood-2003-01-0049
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发表时间：
2003-09-15
期刊：
影响因子：
20.3
通讯作者：
Rafii, S
中科院分区：
文献类型：
--
作者：
Zhang, F;Hackett, NR;Rafii, S
包括绿色荧光蛋白（GFP）在内的报告基因已被用于监测基因转移载体导入血管细胞的转基因表达。在这里，我们证明了GFP本身可以选择性地诱导内皮细胞中某些基因的表达。内皮细胞中GFP的细胞质浓度升高，导致热休克蛋白70 （HSP70）的显著上调。GFP诱导HSP70 mRNA和蛋白表达呈剂量依赖性。gfp介导的HSP70上调导致环氧化酶-2 （COX-2）的诱导，随后前列腺素E-2 （PGE(2)）的产生。gfp介导的HSP70的上调不依赖于丝裂原活化蛋白激酶和磷脂酰肌醇-3激酶信号级联，因为抑制这些途径对HSP70的升高没有影响。腺病毒将GFP传递到小鼠血管中，可显著增强血流量，表明产生了足够的PGE2来诱导血管舒张。鉴定与GFP相互作用的分子伙伴将增加我们对调节应激血管生成和止血的血管特异性因子的理解。
Reporter genes, including green fluorescent protein (GFP), have been used to monitor the expression of transgenes introduced into vascular cells by gene transfer vectors. Here, we demonstrate that GFP by itself can selectively induce expression of certain genes in endothelial cells. Elevation of the cytoplasmic concentration of GFP in endothelial cells, specifically, resulted in a robust upregulation of heat shock protein 70 (HSP70). GFP induced both mRNA and protein expression of HSP70 in a dose-dependent manner. GFP-mediated upregulation of HSP70 resulted in induction of cyclooxygenase-2 (COX-2) followed by prostaglandin E-2 (PGE(2)) production. GFP-mediated up-regulation of HSP70 is independent of mitogen-activated protein kinase and phosphatidylinositol-3-kinase signaling cascades because inhibition of these pathways had no effect on HSP70 increases. Adenoviral delivery of GFP into murine vasculature significantly enhanced blood flow, suggesting that sufficient PGE2 is produced to induce vasodilation. Identification of the molecular partners that interact with GFP will increase our understanding of the vascular-specific factors that regulate stress angiogenesis and hemostasis.