The influence of YS-1 on the Dll4-Notch1 signaling pathway.
The influence of YS-1 on the Dll4-Notch1 signaling pathway.
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DOI：
10.1093/abbs/gmt125
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发表时间：
2014
影响因子：
3.7
通讯作者：
Li Sun;Qingqing Yang;Ping Wang;Datao Liu;Wen-lu Liang;Sen-sen Lin;S. Yuan
中科院分区：
文献类型：
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作者：
Li Sun;Qingqing Yang;Ping Wang;Datao Liu;Wen-lu Liang;Sen-sen Lin;S. Yuan
本研究旨在探讨p43和YS-1（重组人p43蛋白）在Dll 4-Notch 1信号通路中的作用及其分子机制。以p43蛋白为靶点，用活性小干扰RNA和重组质粒感染人脐静脉内皮细胞（HUVECs）。采用三维出芽模型、内皮细胞迁移实验、出芽和管腔形成实验等方法研究p43和YS-1在血管生成中的作用。采用半定量逆转录聚合酶链反应（RT-PCR）和蛋白质印迹法（Western blot）检测p43在Dll 4-Notch 1信号转导中的作用。研究发现，p43的沉默和过表达可上调Dll 4-Notch并刺激血管生成。p43在血管生成中起着复杂的作用。当浓度低于100 nM时，它促进血管生成;相反，当浓度超过100 nM时，它抑制血管生成。在本研究中，我们发现p43的表达水平低于60 nM。然而，重组人p43蛋白YS-1抑制内皮细胞发芽，并且500 μg/ml YS-1减弱Dll 4-Notch 1信号传导的激活。提示YS-1可通过Dll 4-Notch 1信号转导途径直接抑制血管生成，而p43在该信号转导途径中起调节作用。
In this study, we investigated the role and molecular mechanism of p43 and YS-1 (recombinant human p43 protein) in Dll4-Notch1 signaling pathway. Active, small interfering RNA and recombinant plasmid targeting of p43 protein were used to infect human umbilical vein endothelial cells (HUVECs). Three-dimensional sprouting model, endothelial cell migration assay, and sprouting and tube formation assay were used to deduce the function of p43 and YS-1 in angiogenesis. Semi-quantitative reverse transcription-polymerase chain reaction and western blot analysis were performed to detect the efficiency of p43 in Dll4-Notch1 signaling in HUVECs. It was found that silencing and overexpression of p43 could upregulate Dll4-Notch and stimulate angiogenesis. p43 plays a complex role in angiogenesis. When the concentration is under 100 nM, it promotes angiogenesis; instead, when the concentration is over 100 nM, it inhibits angiogenesis. In this study, we found that the expression level of p43 was under 60 nM. However, recombinant human p43 protein, YS-1, inhibited endothelial cell sprouting, and 500 μg/ml of YS-1 attenuated the activation of Dll4-Notch1 signaling. These results suggested that YS-1 could directly inhibit angiogenesis through Dll4-Notch1 signal transduction pathway, while p43 plays a modulating role in this signaling pathway.