Impact of endostatin on bFGF-induced proliferation, migration, and matrix metalloproteinase-2 expression/secretion of bovine choroidal endothelial cells.
OBJECTIVE
To investigate the potential role of endostatin, an endogenous angiogenesis inhibitor, in the prevention of choroidal angiogenesis-related disorders.


METHODS
Bovine choroidal endothelial cells (CEC) were cultured and treated with basic fibroblast growth factor (bFGF) alone or combined with endostatin at concentrations ranging from 0.1 to 10 microg/ml. The proliferation and migration of CECs were evaluated by using 3, (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay and modified Boyden chamber assay, respectively. For evaluating expression and secretion of matrix metalloproteinase-2 (MMP-2), CEC-conditioned media were subjected to zymography and/or Western blot analysis, and the cells were used for semiquantitative reverse transcription polymerase chain reaction (RT-PCR) analysis.


RESULTS
Endostatin did not inhibit bFGF-induced or nonstimulated CEC proliferation (p > 0.05). The bFGF-induced migration was significantly inhibited by endostatin at concentrations of 1 and 10 microg/ml (p < 0.05). The bFGF-upregulated expression of mRNA in CECs and the secretion of MMP-2 protein of CECs were both suppressed by endostatin.


CONCLUSIONS
Inhibitory effect of endostatin on expression and secretion of MMP-2 and cell migration, but not on proliferation of CECs, could respond to its therapeutic action for choroidal neovascularization-dependent disorders.
Wang
Yu-Sheng
YS
Department of Ophthalmology, Faculty of Medicine, University of Leipzig, Liebigstrasse 10-14, 04103 Leipzig, Germany.
Eichler
Wolfram
W
Friedrichs
Ulrike
U
Yafai
Yousef
Y
Hoffmann
Stephan
S
Yasukawa
Tsutomu
T
Hui
Yan-Nian
YN
Wiedemann
Peter
P
D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002829:Choroid / Q000098:blood supply*; D015536:Down-Regulation; D043169:Endostatins / Q000494:pharmacology*; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism*; D061965:Matrix Metalloproteinase Inhibitors; D012333:RNA, Messenger / Q000378:metabolism
D020533:Angiogenesis Inhibitors; D043169:Endostatins; D061965:Matrix Metalloproteinase Inhibitors; D012333:RNA, Messenger; D016222:Fibroblast Growth Factor 2; D020778:Matrix Metalloproteinase 2