c-jun N-terminal kinase activation by hydrogen peroxide in endothelial cells involves Src-dependent epidermal growth factor receptor transactivation
c-jun N-terminal kinase activation by hydrogen peroxide in endothelial cells involves Src-dependent epidermal growth factor receptor transactivation
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DOI：
10.1074/jbc.m011766200
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发表时间：
2001-05-11
影响因子：
4.8
通讯作者：
Keaney, JF
中科院分区：
文献类型：
--
作者：
Chen, K;Vita, JA;Keaney, JF
内皮细胞的表型特性受到氧化应激的调节，并且c-dun N-末端激酶（JNK）途径在介导细胞对应激的反应中是重要的，尽管该途径在内皮细胞中的活化尚未完全表征。因此，我们将内皮细胞暴露于过氧化氢（H2O2），并观察到JNK在15分钟内快速活化，其涉及JNK和c-Jun的磷酸化以及AP-1 DNA结合活性的诱导，蛋白激酶C和磷酸肌醇3-激酶的抑制不影响JNK活化。与此相反，酪氨酸激酶抑制剂染料木黄酮、除草霉素A和4-氨基-5-氨基-2-甲基-N-甲基-N，（4-氯苯基）-7-（叔丁基）吡唑并[3，4-D]嘧啶（PP2）显著减弱H2O2诱导的JNK活化，用显性阴性形式的Src转染内皮细胞腺病毒也是如此，暗示Src是JNK的上游活化剂，AG1478和针对表皮生长因子受体（EGFR）的反义寡核苷酸也抑制了H2O2对JNK的激活，表明EGFR参与了这一过程。与此观察结果一致，H2O2刺激EGFR酪氨酸磷酸化和与Shc-Grb2形成复合物，而PP2则消除了该复合物，这表明Src参与了H2O2诱导的EGFR活化。通过自磷酸化特异性抗体评估，H2O2对EGFR的酪氨酸磷酸化不涉及Tyr（1173）处的受体自磷酸化。这些数据表明，H2O2诱导的JNK激活内皮细胞涉及EGFR通过Src依赖性途径，这是从EGFR配体激活不同。这些数据代表了一个潜在的途径，介导氧化应激诱导的内皮细胞表型变化。
The phenotypic properties of the endothelium are subject to modulation by oxidative stress, and the c-dun N-terminal kinase (JNK) pathway is important in mediating cellular responses to stress, although activation of this pathway in endothelial cells has not been fully characterized. Therefore, we exposed endothelial cells to hydrogen peroxide (H2O2) and observed rapid activation of JNK within 15 min that involved phosphorylation of JNK and c-Jun and induction of AP-1 DNA binding activity, Inhibition of protein kinase C and phosphoinositide 3-kinase did not effect JNK activation. In contrast, the tyrosine kinase inhibitors, genistein, herbimycin A, and 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-D]pyrimidine (PP2) significantly attenuated H2O2-induced JNK activation as did endothelial cell adenoviral transfection with a dominant-negative form of Src, implicating Src as an upstream activator of JNK, Activation of JNK by H2O2 was also inhibited by AG1478 and antisense oligonucleotides directed against the epidermal growth factor receptor (EGFR), implicating the EGFR in this process. Consistent with this observation, H2O2 stimulated EGFR tyrosine phosphorylation and complex formation with Shc-Grb2 that was abolished by PP2, implicating Src in H2O2-induced EGFR activation. Tyrosine phosphorylation of the EGFR by H2O2 did not involve receptor autophosphorylation at Tyr(1173) as assessed by an autophosphorylation-specific antibody. These data indicate that H2O2-induced JNK activation in endothelial cells involves the EGFR through an Src-dependent pathway that is distinct from EGFR ligand activation. These data represent one potential pathway for mediating oxidative stress-induced phenotypic changes in the endothelium.