Erbin plays a critical role in human umbilical vein endothelial cell migration and tubular structure formation via the Smad1/5 pathway
Erbin plays a critical role in human umbilical vein endothelial cell migration and tubular structure formation via the Smad1/5 pathway
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Erbin通过Smad1/5途径在人脐静脉内皮细胞迁移和管状结构形成中发挥关键作用
DOI：
10.1002/jcb.27754
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发表时间：
2019-03-01
影响因子：
4
通讯作者：
Guo, Yuan
中科院分区：
文献类型：
--
作者：
Jin, Xiaodong;Li, Bo;Guo, Yuan
血管生成是动脉粥样硬化的重要过程。 ErbB2已被证明在血管发育中具有重要作用，但Erbin是否在血管中表达以及其在血管中的定位和功能仍不清楚。在当前的研究中，我们研究了 Erbin 在人脐静脉中的位置和功能。制备人脐静脉，进行免疫荧光分析以确定Erbin的表达。培养人脐静脉内皮细胞（HUVEC）并制备含有Erbin RNAi的慢病毒（LV）。慢病毒转染后，分别采用CCK-8法和Annexin V-PI法检测细胞增殖和凋亡。使用划痕伤口愈合测定和transwell测定研究细胞迁移。进行毛细管样管形成实验以说明 Erbin 对 HUVEC 管形成的影响。用蛋白质印迹评估信号通路分子的表达。免疫荧光分析表明，Erbin 在人脐静脉中表达，并且大部分 Erbin 在内皮细胞中强烈共定位。虽然Erbin的敲低不影响HUVEC增殖和凋亡，但它显着抑制HUVEC迁移和管状结构形成。 Erbin 敲低对 ERK1/2 和 Smad2/3 信号通路没有影响，但显着促进 Smad1/5 磷酸化和核转位。 Smad1/5 通路的消除降低了 Erbin 对内皮细胞的影响。 Erbin 主要定位于人脐静脉内皮细胞中，通过 Smad1/5 通路在内皮细胞迁移和肾小管形成中发挥关键作用。
Angiogenesis is an important process in atherosclerosis. ErbB2 was proved to have an important role in vascular development, but it is still unclear whether Erbin expresses in vessels as well as its location and function in the vessels. In the current study, we investigated the location and function of Erbin in human umbilical veins. The human umbilical veins were prepared, and immunofluorescent analysis was performed to determine the expression of Erbin. Human umbilical vein endothelial cells (HUVECs) were cultured and the lentivirus (LV) containing Erbin RNAi was also prepared. After transfection with the lentivirus, CCK‐8 assay and Annexin V‐PI assay were used for cell proliferation and apoptosis, respectively. Cell migration was studied using the scratch wound healing assay and the transwell assay. The capillary‐like tube formation assay was performed to illustrate the effect of Erbin on HUVEC tube formation. Expression of signaling pathway molecules was assessed with Western blot. The immunofluorescent analysis suggested that Erbin expressed in human umbilical veins and the majority of the Erbin is strongly colocalized in endothelial cells. Although knockdown of Erbin did not affect HUVEC proliferation and apoptosis, it significantly suppressed HUVEC migration and tubular structure formation. Erbin knockdown showed no effect on the ERK1/2 and Smad2/3 signaling pathways but significantly promoted Smad1/5 phosphorylation and nuclear translocation. Ablation of the Smad1/5 pathway decreased the effects of Erbin on endothelial cells. Erbin is mainly localized in endothelial cells in human umbilical veins and plays a critical role in endothelial cell migration and tubular formation via the Smad1/5 pathway.