Department of Cardiology, Qilu Hospital, Shandong University, Jinan, Shandong, China.
The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Shandong University, Jinan, Shandong, China.
J Cell Biochem. 2019 Mar;120(3):4654-4664. doi: 10.1002/jcb.27754. Epub 2018 Oct 26.
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.
血管生成是动脉粥样硬化的一个重要过程。ErbB2 被证明在血管发育中具有重要作用,但它在血管中的表达及其位置和功能仍不清楚。在本研究中,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)。转染慢病毒后,通过 CCK-8 assay 和 Annexin V-PI assay 分别用于细胞增殖和凋亡分析。通过划痕愈合试验和 Transwell 试验研究细胞迁移。通过毛细血管样管形成试验说明 Erbin 对 HUVEC 管形成的影响。通过 Western blot 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 强烈共定位于内皮细胞中。虽然 Erbin 的敲低不影响 HUVEC 的增殖和凋亡,但它显著抑制 HUVEC 的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,但显著促进 Smad1/5 的磷酸化和核转位。Smad1/5 通路的阻断降低了 Erbin 对内皮细胞的影响。Erbin 主要定位于人脐静脉的内皮细胞中,通过 Smad1/5 通路在内皮细胞迁移和管状形成中发挥关键作用。