Weisen Wang1,2, Zhi Wang1, Dingyuan Tian1,2, Xi Zeng1, Yangdong Liu3, Qining Fu3, Anlin Liang4, Yi Zhang1, Qiangguo Gao1, Jizhong Cheng5, Yun Wang1. 1. Department of Cell Biology, College of Basic Medical Sciences, Army Medical University (Third Military Medical University), Chongqing, China. 2. Cadet Battalion, College of Basic Medical Sciences, Army Medical University (Third Military Medical University), Chongqing, China. 3. Department of Vascular Surgery, First Affiliated Hospital of Chongqing Medical University, Chongqing, China. 4. Department of Orthopaedics, First Affiliated Hospital of Chongqing Medical University, Chongqing, China. 5. Department of Medicine, Baylor College of Medicine, Houston, Texas, USA.
Abstract
BACKGROUND/AIMS: Neointimal hyperplasia is responsible for stenosis, which requires corrective vascular surgery, and is also a major morphological feature of many cardiovascular diseases. This hyperplasia involves the endothelial-to-mesenchymal transition (EndMT). We investigated whether integrin β3 can modulate the EndMT, as well as its underlying mechanism. METHODS: Integrin β3 was overexpressed or knocked down in human umbilical vein endothelial cells (HUVECs). The expression of endothelial markers and mesenchymal markers was determined by real-time reverse transcription PCR (RT-PCR), immunofluorescence staining, and western blot analysis. Notch signaling pathway components were detected by real-time RT-PCR and western blot analysis. Cell mobility was evaluated by wound-healing, Transwell, and spreading assays. Fibroblast-specific protein 1 (FSP-1) promoter activity was determined by luciferase assay. RESULTS: Transforming growth factor (TGF)-β1 treatment or integrin β3 overexpression significantly promoted the EndMT by downregulating VE-cadherin and CD31 and upregulating smooth muscle actin α and FSP-1 in HUVECs, and by enhancing cell migration. Knockdown of integrin β3 reversed these effects. Notch signaling was activated after TGF-β1 treatment of HUVECs. Knockdown of integrin β3 suppressed TGF-β1-induced Notch activation and expression of the Notch downstream target FSP-1. CONCLUSION: Integrin β3 may promote the EndMT in HUVECs through activation of the Notch signaling pathway.
BACKGROUND/AIMS: Neointimal hyperplasia is responsible for stenosis, which requires corrective vascular surgery, and is also a major morphological feature of many cardiovascular diseases. This hyperplasia involves the endothelial-to-mesenchymal transition (EndMT). We investigated whether integrin β3 can modulate the EndMT, as well as its underlying mechanism. METHODS: Integrin β3 was overexpressed or knocked down in human umbilical vein endothelial cells (HUVECs). The expression of endothelial markers and mesenchymal markers was determined by real-time reverse transcription PCR (RT-PCR), immunofluorescence staining, and western blot analysis. Notch signaling pathway components were detected by real-time RT-PCR and western blot analysis. Cell mobility was evaluated by wound-healing, Transwell, and spreading assays. Fibroblast-specific protein 1 (FSP-1) promoter activity was determined by luciferase assay. RESULTS: Transforming growth factor (TGF)-β1 treatment or integrin β3 overexpression significantly promoted the EndMT by downregulating VE-cadherin and CD31 and upregulating smooth muscle actin α and FSP-1 in HUVECs, and by enhancing cell migration. Knockdown of integrin β3 reversed these effects. Notch signaling was activated after TGF-β1 treatment of HUVECs. Knockdown of integrin β3 suppressed TGF-β1-induced Notch activation and expression of the Notch downstream target FSP-1. CONCLUSION: Integrin β3 may promote the EndMT in HUVECs through activation of the Notch signaling pathway.
Authors: Boyd R Rorabaugh; Nathaniel W Mabe; Sarah L Seeley; Thorne S Stoops; Kasey E Mucher; Connor P Ney; Cassandra S Goodman; Brooke J Hertenstein; Austen E Rush; Charis D Kasler; Aaron M Sargeant; Phillip R Zoladz Journal: Stress Date: 2019-07-26 Impact factor: 3.493
Authors: Timothy E Gressett; Danielle Nader; Juan Pablo Robles; Tione Buranda; Steven W Kerrigan; Gregory Bix Journal: Front Cell Infect Microbiol Date: 2022-04-29 Impact factor: 6.073