title: A C-terminal fragment BIGH3 protein with an RGDRGD motif inhibits corneal neovascularization in vitro and in vivo.
abstract: An Arg-Gly-Asp (RGD) motif in the fourth FAS1 domain of the human BIGH3 (transforming growth factor-β1-inducible gene-h3) protein has been reported to play an important role in mediating tumor angiogenesis. The aim of this study was to investigate the inhibitory effect of a modified C-terminal fragment BIGH3 protein with an RGDRGD motif on corneal neovascularization in vitro and in vivo. Recombinant C-terminal fragment BIGH3 protein with wild-type sequence and modified C-terminal fragment BIGH3 protein containing an RGDRGD motif were successfully expressed and purified. We demonstrated that both proteins significantly inhibited vascular endothelial growth factor (VEGF)-induced human umbilical vein endothelial cell (HUVEC) adhesion, migration, and tube formation and induced cell apoptosis but failed to inhibit HUVEC proliferation. We determined that the mechanism underlying this activity was an interaction between BIGH3 and αvβ3 integrin, which blocked the phosphorylation of PI3K/Akt and ERK signaling pathways. The inhibitory effects of wild-type and modified C-terminal fragment BIGH3 proteins on angiogenesis were confirmed by a rabbit corneal neovascularization assay. More importantly, we provided evidence that the modified C-terminal fragment BIGH3 protein with an RGDRGD motif inhibited angiogenic activity far more effectively than did wild-type C-terminal fragment BIGH3. Collectively, our data show that a C-terminal fragment BIGH3 protein containing an RGDRGD motif might be promising as an effective drug in treating corneal neovascularization.
authors.affiliation: Key Laboratory of Harbin Medical University Eye Center, Eye Hospital, First Affiliated Hospital, Harbin Medical University, 23 Youzheng Street, Nangang District, Harbin 150001, PR China.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Ge
authors.forename: Pei
authors.initials: P
authors.lastname: Tian
authors.forename: Linan
authors.initials: L
authors.lastname: Guan
authors.forename: Xiuli
authors.initials: X
authors.lastname: Yin
authors.forename: Hanruo
authors.initials: H
authors.lastname: Liu
authors.forename: Nan
authors.initials: N
authors.lastname: Xiao
authors.forename: Yongzhong
authors.initials: Y
authors.lastname: Xiong
authors.forename: Xin
authors.initials: X
authors.lastname: Luo
authors.forename: Yunduan
authors.initials: Y
authors.lastname: Sun
authors.forename: Donghua
authors.initials: D
authors.lastname: Qi
authors.forename: Shuang
authors.initials: S
authors.lastname: Ni
authors.forename: Ping
authors.initials: P
authors.lastname: Liu
mesh_terms: D020816:Amino Acid Motifs; D020533:Angiogenesis Inhibitors / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001808:Blood Vessels / Q000187:drug effects; D015153:Blotting, Western; D020169:Caspases / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D016510:Corneal Neovascularization / Q000378:metabolism / Q000517:prevention & control*; D004195:Disease Models, Animal; D016326:Extracellular Matrix Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D005434:Flow Cytometry; D061307:Human Umbilical Vein Endothelial Cells; D039302:Integrin alphaVbeta3 / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation; D010957:Plasmids; D017510:Protein Folding; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D011817:Rabbits; D016212:Transforming Growth Factor beta / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D000099222:betaIG-H3 Protein
chemical_list: D020533:Angiogenesis Inhibitors; D020169:Caspases; D016326:Extracellular Matrix Proteins; D039302:Integrin alphaVbeta3; D009842:Oligopeptides; D010446:Peptide Fragments; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D016212:Transforming Growth Factor beta; D042461:Vascular Endothelial Growth Factor A; D000099222:betaIG-H3 Protein; C047981:arginyl-glycyl-aspartic acid