Background: This study aimed to investigate the anti-angiogenesis effect of 7,8-dihydroxyflavone (7,8-DHF) and its potential molecular mechanism. Methods: The rhesus macaque choroid-retinal endothelial (RF/6A) cells were treated with different concentrations (from 0 to 100 ÎŒM) of 7,8-DHF and/or 40 ng/ml VEGF. The morphology, proliferation, migration, capillary-like tube formationïŒand apoptosis of RF/6A cells were evaluated by Giemsa staining, CCK-8 assay, transwell migration assay, matrigel tube formation assay, and flow cytometry/hoechst33342 staining, respectively. The protein content of VEFGR2 and p-VEGFR2 was assessed by western blotting. Results: 7,8-DHF significantly inhibited the proliferation, migration, and tube formation of RF/6A cells and promoted their apoptosis in vitro. The expression of VEGFR2 in RF/6A cells was constant whether or not to administer 7,8-DHF. However, the phosphorylation of VEGFR2 significantly decreased after the administration of 7,8-DHF. Conclusions: 7,8-DHF could inhibit RF/6A angiogenesis in vitro. The inhibitory mechanism of 7,8-DHF in angiogenesis was attributed to the suppression of VEGFR2 phosphorylation and thus blocking of VEGF/VEGFR2 signal pathway.