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Erbin plays a critical role in human umbilical vein endothelial cell migration and tubular structure formation via the Smad1/5 pathway
Authors:Xiaodong Jin  Bo Li  Yunhe Zhao  Xiqiang Liu  Yuhua Li  Lina Song  Lifang Cui  Dan Xie  Tao Li  Xiufang Zhang  Yuan Guo
Institution:1. 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

Department of Geriatrics, Central Hospital of Zibo, Zibo, China;2. Heart Center, Central Hospital of Zibo, Zibo, China;3. Department of Geriatrics, Central Hospital of Zibo, Zibo, China;4. Department of CT and Magnetic Resonance, Central Hospital of Zibo, Zibo, China;5. Department of Hospital Office, Maternal and Child Health Care Hospital of Zibo, Zibo, China;6. Department of Hyperbaric Oxygen, Central Hospital of Zibo, Zibo, China;7. Department of Rehabilitation Medicine, Sixth People's Hospital of Zibo, Zibo, China;8. Center of Translational Medicine, Central Hospital of Zibo, Zibo, China;9. Department of Medical Administration Division, Central Hospital of Zibo, Zibo, China;10. Department of Cardiology, Qilu Hospital, Shandong University, Jinan, Shandong, China

Abstract: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.
Keywords:Erbin  human umbilical vein endothelial cells (HUVECs)  migration  Smad1/5  tubular structure formation
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