title: Nestin-mediated cytoskeletal remodeling in endothelial cells: novel mechanistic insight into VEGF-induced cell migration in angiogenesis.
abstract: Nestin is highly expressed in poorly differentiated and newly formed proliferating endothelial cells (ECs); however, the role of this protein in angiogenesis remains unknown. Additionally, the cytoskeleton and associated cytoskeleton-binding proteins mediate the migration of vascular ECs. Therefore, the aim of the present study was to determine whether VEGF regulates the cytoskeleton, as well as other associated proteins, to promote the migration of vascular ECs. The coexpression of nestin and CD31 during angiogenesis in alkali-burned rat corneas was examined via immunohistochemical analysis. Western blot analyses revealed that the exposure of human umbilical vein endothelial cells (HUVECs) to hypoxia promoted nestin expression in vitro. Additionally, nestin silencing via siRNA significantly inhibited many of the process associated with VEGF-induced angiogenesis, including tube formation and the migration and proliferation of HUVECs. Moreover, FITC-phalloidin labeling revealed that F-actin filaments were successfully organized into microfilaments in VEGF-treated cells, suggesting a network rearrangement accomplished via F-actin that contrasted with the uniform and loose actin filament network observed in the siRNA-nestin cells. The results of the present study highlight the key role played by nestin in activated HUVECs during angiogenesis. The inhibition of the ERK pathway suppressed the nestin expression induced by VEGF in the HUVECs. Therefore, our study provides the first evidence that nestin-mediated cytoskeleton remodeling in ECs occurs via filopodia formation along the cell edge, facilitating both filopodia localization and cell polarization and ultimately promoting HUVEC migration via VEGF induction, which may be associated with ERK pathway activation.
authors.affiliation: Department of Biochemistry, Zhongshan Medical School, Sun Yat-sen University, Guangzhou, Guangdong Province, China;
authors.forename: Zhen-Wei
authors.initials: ZW
authors.lastname: Liang
authors.affiliation: Department of Biochemistry, Zhongshan Medical School, Sun Yat-sen University, Guangzhou, Guangdong Province, China;
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Obstetrics and Gynecology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China; Center for Disease Model Animals, Sun Yat-sen University, Guangzhou, China, Guangzhou, Guangdong Province, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Biochemistry, Zhongshan Medical School, Sun Yat-sen University, Guangzhou, Guangdong Province, China;
authors.forename: Cen
authors.initials: C
authors.lastname: Li
authors.affiliation: Department of Biochemistry, Zhongshan Medical School, Sun Yat-sen University, Guangzhou, Guangdong Province, China;
authors.forename: Ti
authors.initials: T
authors.lastname: Zhou
authors.affiliation: Department of Biochemistry, Zhongshan Medical School, Sun Yat-sen University, Guangzhou, Guangdong Province, China;
authors.forename: Zhonghan
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Department of Biochemistry, Zhongshan Medical School, Sun Yat-sen University, Guangzhou, Guangdong Province, China;
authors.forename: Xia
authors.initials: X
authors.lastname: Yang
authors.affiliation: Department of Biochemistry, Zhongshan Medical School, Sun Yat-sen University, Guangzhou, Guangdong Province, China;
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Department of Biochemistry, Zhongshan Medical School, Sun Yat-sen University, Guangzhou, Guangdong Province, China; Key Laboratory of Functional Molecules from Marine Microorganisms (Sun Yat-sen University), Department of Education of Guangdong Province, Guangzhou, Guangdong Province, China; and.
authors.forename: Guoquan
authors.initials: G
authors.lastname: Gao
authors.affiliation: Department of Biochemistry, Zhongshan Medical School, Sun Yat-sen University, Guangzhou, Guangdong Province, China; Center for Disease Model Animals, Sun Yat-sen University, Guangzhou, China, Guangzhou, Guangdong Province, China caiwb@mail.sysu.edu.cn gaogq@mail.sysu.edu.cn.
authors.forename: Weibin
authors.initials: W
authors.lastname: Cai
mesh_terms: D043925:Angiogenesis Inducing Agents / Q000494:pharmacology*; D000818:Animals; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000187:drug effects / Q000502:physiology*; D042783:Endothelial Cells / Q000187:drug effects / Q000502:physiology*; D005260:Female; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000502:physiology; D006801:Humans; D064231:Nestin / Q000502:physiology*; D011554:Pseudopodia / Q000187:drug effects / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology*
chemical_list: D043925:Angiogenesis Inducing Agents; C577484:NES protein, human; D064231:Nestin; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A
keywords: VEGF; angiogenesis; cytoskeleton; nestin