title: Novel bioactivity of NHERF1 in corneal neovascularization.
abstract: OBJECTIVE
  This paper aimed to determine the involvement of Na(+)/H(+) exchanger regulatory factor 1 (NHERF1) in experimental murine corneal neovascularization (NV), and to study the effect of NHERF1 on the biological properties of HUVEC and related mechanisms.
  
  
  METHODS
  Using loss- and gain-function, we investigated the biological effects of NHERF1 on HUVEC. Western blotting was used to detect the expression of NHERF1 in cells. A carboxyfluorescein succinimidyl ester (CFSE) labeling assay and scarification test were used to measure the proliferation and migration activity, respectively, of HUVEC. The cell cycle distribution of the cells was assessed by flow cytometry analysis. The effect of NHERF1 on the phosphorylation levels of Akt and the changes of matrix metalloproteinase (MMP)-2 and MMP-9 levels were detected by western blotting analysis. Change in the NHERF1 expression in the alkali burn-induced corneal NV model was detected by microarray, real-time PCR, and immunofluorescence.
  
  
  RESULTS
  Overexpression of exogenous NHERF1 in HUVEC slightly inhibited the proliferation and significantly reduced the migration of the cells. NHERF1 also significantly downregulated Akt1 phosphorylation induced by platelet-derived growth factor BB (PDGF-BB) and the secretion of MMP-2 and MMP-9 compared with control cells. NHERF1 was upregulated in corneas challenged with alkali burns.
  
  
  CONCLUSIONS
  Our results indicated that NHERF1 might serve as a potential target for manipulating neovascularization-related diseases. This discovery contributes to a better understanding of the bioactivity of NHERF1 in angiogenesis.
authors: State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of Medical Sciences, Qingdao, China.
authors: Peng
authors: P
authors: Chen
authors: Ye
authors: Y
authors: Wang
authors: Lingling
authors: L
authors: Yang
authors: Changyou
authors: C
authors: Li
authors: Yao
authors: Y
authors: Wang
authors: Lixin
authors: L
authors: Xie
authors: Yiqiang
authors: Y
authors: Wang
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002057:Burns, Chemical / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D016510:Corneal Neovascularization / Q000378:metabolism* / Q000473:pathology; D003320:Corneal Ulcer / Q000378:metabolism; D005126:Eye Burns / Q000139:chemically induced; D005260:Female; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000201:enzymology; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D010750:Phosphoproteins / Q000494:pharmacology / Q000502:physiology*; D010766:Phosphorylation; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction; D012972:Sodium Hydroxide; D017923:Sodium-Hydrogen Exchangers / Q000494:pharmacology / Q000502:physiology*; D014162:Transfection
chemical_list: D000199:Actins; D010750:Phosphoproteins; D010982:Platelet-Derived Growth Factor; D017923:Sodium-Hydrogen Exchangers; C093376:sodium-hydrogen exchanger regulatory factor; D012972:Sodium Hydroxide; C514862:Akt1 protein, mouse; D051057:Proto-Oncogene Proteins c-akt; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9