Mitogenic and antiapoptotic actions of hepatocyte growth factor through ERK, STAT3, and Akt in endothelial cells
Mitogenic and antiapoptotic actions of hepatocyte growth factor through ERK, STAT3, and Akt in endothelial cells
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DOI：
10.1161/01.hyp.37.2.581
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发表时间：
2001-02-01
期刊：
影响因子：
8.3
通讯作者：
Ogihara, T
中科院分区：
文献类型：
--
作者：
Nakagami, H;Morishita, R;Ogihara, T
肝细胞生长因子（HGF）是血管生成生长因子的一员，在内皮细胞的调节中可能起关键作用，因为HGF在内皮细胞中显示促有丝分裂和抗凋亡作用。由于这些作用的机制尚不清楚，我们研究了HGF在人主动脉内皮细胞中的信号转导系统。用重组肝细胞生长因子（rHGF）治疗内皮细胞导致DNA合成显著增加，如胸苷掺入所评估。重要的是，清楚地观察到rHGF对细胞外信号相关激酶（ERK）和Akt的磷酸化。因此，我们进一步研究了ERK或Akt的特异性抑制剂对细胞增殖的影响。PD 98059（一种促分裂原活化蛋白激酶激酶抑制剂）预处理可显著减弱rHGF诱导的细胞增殖，而磷脂酰肌醇-3-OH激酶抑制剂渥曼青霉素和LY-294002则无此作用。有趣的是，用rHGF处理显著增加了信号转导和转录激活因子（STAT）3（Ser 727）的磷酸化，而PD 98059减弱了由rHGF诱导的Ser 727的磷酸化。此外，用rHGF处理显著增加了c-Sos的启动子活性，c-Sos包括sis诱导元件和血清反应元件，而PD 98059完全减弱rHGF诱导的c-fos启动子的激活。相反，渥曼青霉素和LY-294002抑制Akt不能抑制STAT 3磷酸化和c-fos激活。另一方面，rHGF处理减弱了由肿瘤坏死因子-α刺激引起的LDH释放和caspase-3活性的增加。与DNA合成相反，wortmannin和LY-294002显著减弱了由rHGF介导的caspase-3活性的降低，而PD 98059则没有。总之，本研究表明，肝细胞生长因子刺激细胞增殖通过ERK-STAT 3（Ser 727）途径，并通过磷脂酰肌醇-3-OH激酶-Akt途径在人主动脉内皮细胞的抗凋亡作用。这些发现为HGF在心血管疾病中的作用提供了新的视角。
Hepatocyte growth factor (HGF), a member of the angiogenic growth factors, may play a pivotal role in the regulation of endothelial cells, inasmuch as HGF shows mitogenic and antiapoptotic actions in endothelial cells. Because the mechanism of these actions is still unclear, we examined the signal transduction system of HGF in human aortic endothelial cells. Treatment of endothelial cells with recombinant HGF (rHGF) resulted in a significant increase in DNA synthesis as assessed by thymidine incorporation. Importantly, phosphorylation of extracellular signal-related kinase (ERK) and Akt by rHGF was clearly observed. Thus, we further examined the effects of specific inhibitors of ERK or Akt on cell proliferation. Pretreatment with PD98059, a mitogen-activated protein kinase kinase inhibitor, significantly attenuated cell proliferation induced by rHGF, whereas inhibitors of phosphatidylinositol-3-OH kinase, wortmannin, and LY-294002, did not. Interestingly, treatment with rHGF significantly increased the phosphorylation of the signal transducers and activators of transcription (STAT)3 (Ser727), whereas PD98059 attenuated the phosphorylation of Ser727 induced by rHGF, In addition, treatment with rHGF significantly increased the promoter activity of c-Sos, which includes the sis-inducible element and serum response element, whereas PD98059 completely attenuated the activation of the c-fos promoter induced by rHGF. In contrast, inhibition of Akt by wortmannin and LY-294002 failed to inhibit the phosphorylation of STAT3 and c-fos activation. On the other hand, treatment with rHGF attenuated the increase in LDH release and caspase-3 activity induced by tumor necrosis factor-alpha stimulation, In contrast to DNA synthesis, wortmannin and LY-294002 markedly attenuated the decrease in caspase-3 activity mediated by rHGF, whereas PD98059 did not. Overall, the present study demonstrated that HGF stimulated cell proliferation through the ERK-STAT3 (Ser727) pathway and had an antiapoptotic action through the phosphatidylinositol-3-OH kinase-Akt pathway in human aortic endothelial cells. These findings provide new perspectives in the role of HGF in cardiovascular disease.