The 15(S)-hydroxyeicosatetraenoic acid-induced angiogenesis requires Janus kinase 2-signal transducer and activator of transcription-5B-dependent expression of interleukin-8
The 15(S)-hydroxyeicosatetraenoic acid-induced angiogenesis requires Janus kinase 2-signal transducer and activator of transcription-5B-dependent expression of interleukin-8
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DOI：
10.1182/blood-2008-10-183210
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发表时间：
2009-06-04
期刊：
影响因子：
20.3
通讯作者：
Rao, Gadiparthi N.
中科院分区：
文献类型：
--
作者：
Cheranov, Sergey Y.;Wang, Dong;Rao, Gadiparthi N.
为了了解15(S)-羟基二糖四烯酸（15(S)-HETE）诱导血管生成的分子基础，我们研究了Janus激酶信号转导和转录激活因子（Jak-STAT）信号的作用。在人视网膜微血管内皮细胞（HRMVECs）中，15(S)-HETE以时间依赖性的方式刺激Jak2的酪氨酸磷酸化。通过腺病毒介导的显性阴性突变体15(S)- hete诱导的HRMVEC迁移、管形成和基质塞血管生成抑制Jak2活化同样，15(S)-HETE以时间依赖性的方式激活STAT-5B的酪氨酸磷酸化。显性负突变介导的STAT-5B激活干扰抑制了15(S)- hete诱导的HRMVEC迁移、管形成和Matrigel塞血管生成。15(S)-HETE以jak2 - stat - 5b依赖性方式诱导人造血干细胞中白细胞介素-8 （IL-8）的表达。此外，中和抗il -8抗体可减少15(S)- hete诱导的HRMVEC迁移、管形成和Matrigel塞血管生成。IL-8启动子的克隆和Transfac分析显示，在-476 nt处存在1个推测的stat结合序列，电泳迁移率转移试验和染色质免疫沉淀分析显示，STAT-5B在15(S)-HETE的作用下与该位点结合。突变分析表明STAT结合位点对15(S)- hete诱导的IL-8启动子活性至关重要。总之，这些观察结果表明，15(S)- hete诱导的血管生成需要依赖jak2 - stat - 5b的IL-8表达。（血液。2009;113:6023-6033）
To understand the molecular basis underlying 15(S)-hydroxyeicosatetraenoic acid (15(S)-HETE)-induced angiogenesis, we have studied the role of the Janus kinase-signal transducer and activator of transcription (Jak-STAT) signaling. The 15(S)-HETE stimulated tyrosine phosphorylation of Jak2 in a time-dependent manner in human retinal microvascular endothelial cells (HRMVECs). Inhibition of Jak2 activation via adenovirus-mediated expression of its dominant-negative mutant attenuated 15(S)-HETE-induced HRMVEC migration and tube formation and Matrigel plug angiogenesis. Similarly, 15(S)-HETE activated tyrosine phosphorylation of STAT-5B in a time-dependent manner. Dominant-negative mutant-mediated interference of STAT-5B activation suppressed 15(S)-HETE-induced HRMVEC migration and tube formation and Matrigel plug angiogenesis. The 15(S)-HETE induced interleukin-8 (IL-8) expression in Jak2-STAT-5B-dependent manner in HRMVECs. In addition, neutralizing anti-IL-8 antibodies reduced 15(S)-HETE-induced HRMVEC migration and tube formation and Matrigel plug angiogenesis. Cloning and Transfac analysis of IL-8 promoter revealed the presence of 1 putative STAT-binding sequence at -476 nt, and electrophoretic mobility shift assay and chromatin immunoprecipitation analysis showed the binding of STAT-5B to this site in response to 15(S)-HETE. Mutational analysis showed that STAT binding site is essential for 15(S)-HETE-induced IL-8 promoter activity. Together, these observations suggest that 15(S)-HETE-induced angiogenesis requires Jak2-STAT-5B-dependent expression of IL-8. (Blood. 2009; 113: 6023-6033)