INFLUENCE OF ANGIOGENESIS INHIBITORS, ENDOSTATIN AND PF-4, ON LYMPHANGIOGENESIS

Abstract

This study was designed to screen potential safe and effective inhibitors oflymphangiogenesis. Lymphatic endothelial cells from pig thoracic duct were isolated andcultured. The control group, 3 endostatin, and 3 PF-4 experimental groups were tested for effectson proliferation and distance of migration of the cultured cells by two methods (method of thescraping line and MTT assay), and observations by light, confocal, and electron microscopywere also made. Total cells migrating past the scrape line for the endostatin control group was28.6±1.2 (mean ± standard error) and the 3 endostatin experimental groups (50ng/ml,100ng/ml,and 150ng/ml), respectively, were 17.5±0.6,10.5±0.5, and 4.8±0.3 (all p<0.05 cf control).Migration distance for the endostatin control group was 381.7±9.67:m, and the migrationdistance of the 3 endostatin experimental groups, respectively, were 252.9±5.58, 164.5±7.09,and 91.2±7.98:m (all p<0.05). Cell migration number for the PF-4 control group was 28.3±1.0compared with doses of 40ng/ml, 80ng/ml, or 120ng/ml of PF-4, respectively, were 13.6±0.7,9.5±0.6, and 4.6±0.4 (all p<0.05 cf control). Migration distance of cells in PF-4 control groupwas 419.9±5.87:m, and the 3 PF-4 experimental groups, respectively, were 199.2±8.16,152.5±7.28, and 104.2± 6.70:m (all p<0.05 cf control). The MTT assay confirmed that as theconcentrations of endostatin and PF-4 were increased, the inhibitory ef ect was increased. Weconclude that endostatin and PF-4 are able to inhibit the migration and proliferation oflymphatic endothelial cells, and these effects are dose-dependent.