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AIM:To investigate the inhibitory effects of endostatin-vascular endothelial growth inhibitor (VEGI_(151)) recombinantadenoviruses on neovascularization.METHODS:We used recombinant adenoviruses to treathuman vascular endothelial cell line ECV304,humanhepatocellular carcinoma cell line HepG2,and murinefibroblast cell line L929,in order to study the chimeric geneexpression in these cell lines.Chick choriallantic membrane(CAM) model,rabbit inflammatory corneal neovascularization(CNV) model,and liver cancer-bearing nude mice modelwere employed to investigate the negative biological effectof fusion molecules on neovascularization in vivo.RESULTS:Western blot showed that the molecular weightof fusion protein was about 41kD after infection of ECV304,HepG2 and L929 cells with supernatant of AdhENDO-VEGI_(151).The fusion protein showed a specific inhibitory effect onthe proliferation of ECV304 cells,but no inhibitory effecton the growth of HepG2 and L929 cells (F=13112.13,P=0.0001).In the chick choriallantic membrane (CAM)assay,the expressed fusion protein significantly inhibitedneovascularization.Rabbit inflammatory corneal neovasculari-zation (CNV) induced by intrastromal sutures resulted in auniform neovascular response.In this model,directsubconjunctival injection of AdhENDO-VEGI_(151) expressedthe fusion protein in vivo and suppressed the developmentof CNV.Topical application of AdhENDO-VEGI_(151) led to asignificant suppression of CNV (F=1413.11,P=0.0001),as compared with the control group of AdLacZ.Immunohist-ochemical staining showed the fusion protein dominantlyexpressed in corneal epithelium.Compared with thecontrol group of AdLacZ (4075.9±1849.9mm~3),the averagetumor size of group AdhENDO-VEGI_(151) reduced in size(487.7±241.2mm~3) (F=14.80,P=0.0085),with an inhibitionrate of 88.03%.Immunohistochemical staining showed theadenoviruses carried the fusion gene expressed on livercancer cell membrane.MVD decreased more significantlyin treated mice (30.75±3.31%) than in AdLacZ control(50.25±8.65%) (F=17.72,P=0.0056) with an inhibition rateof 39%. CONCLUSION:Fusion protein expressed by recombinantadenoviruses has a significant inhibitory effect on neovasculari-zation.
AIM: To investigate the inhibitory effects of endostatin-vascular endothelial growth inhibitor (VEGI_ (151)) recombinant adenoviruses on neovascularization. METHODS: We used recombinant adenoviruses to treathuman vascular endothelial cell line ECV304, human hepatocellular carcinoma cell line HepG2, and murine fibroblast cell line L929, in order to study the chimeric gene expression in these cell lines. Chick cell membrane (CAM) model, rabbit inflammatory corneal neovascularization (CNV) model, and liver cancer-bearing nude mice model were employed to investigate the negative biological effect of fusion molecules on neovascularization in vivo .RESULTS: Western blot showed that the molecular weight of fusion protein was about 41 kD after infection of ECV304, HepG2 and L929 cells with supernatant of AdhENDO-VEGI_ (151). The fusion protein showed a specific inhibitory effect on the proliferation of ECV304 cells, but no inhibitory effect on the growth of HepG2 and L929 cells (F = 13112.13, P = 0.0001) .In the chick (CAM) assay, the expressed fusion protein significantly inhibited neovascularization. Rabbit inflammatory corneal neovasculari-zation (CNV) induced by intrastromal sutures resulted in a uniform neovascular response.In this model, directsubconjunctival injection of AdhENDO-VEGI_ (151) expressedthe fusion protein in vivo and suppressed the developmentof CNV.Topical application of AdhENDO-VEGI_ (151) led to asignificant suppression of CNV (F = 1413.11, P = 0.0001), as compared with the control group of AdLacZ.Immunohist- ochemical staining showed the fusion protein dominantlyexpressed in a corneal epithelium. Compared to the control group of AdLacZ (4075.9 ± 1849.9 mm ~ 3), the averagetumor size of group AdhENDO-VEGI_151 reduced in size (487.7 ± 241.2 mm ~ 3) (F = 14.80, P = 0.0085) , with an inhibition rate of 88.03%. Immunohistochemical staining showed the adenoviruses carrying the fusion gene expressed on liver cancer cell membrane. MDV decreased more significantly in treated mice (30.75 ± 3.31%) than in AdLacZ control (50.25 ± 8.65%) (F = 17.72, P = 0.0056) with an inhibition rate of 39%. CONCLUSION: Fusion protein expressed by recombinant adenoviruses has a significant inhibitory effect on neovasculariation.