Novel anti-angiogenesis activity of fascaplysin via VEGF blockage was recently revealed by our previous study in addition to the reported cyclin-dependent kinase 4 (CDK4) selective inhibition. To uncover more details of this pharmacologically prospective property, this study further investigated whether fascaplysin had direct anti-proliferation effects on human umbilical vein endothelial cells (HUVEC), which might be contributing to anti-angiogenesis. The results showed that G1 cell cycle arrest was induced by 2.6μM fascaplysin in a time-dependent manner, and exhibited more sensitive than hepatocarcinoma cells BeL-7402 and Hela cells. Approximately 56.09±2.63% of the cells were arrested at the G1 phase after 24h, and 64.94±2.07% after 36h, comparing to the 22.82±1.2% in methanol treated cells. Apoptosis of HUVEC cells was induced by 1.3μM fascaplysin and indicated by the sub-G1, Hoechst staining, terminal deoxynucleotidyl transferase dUTP-mediated nicked end labeling (TUNEL) assay, and annexin-V and propidium (PI) label. This apoptosis response was further confirmed by the detection of active caspase-3 and by western blotting using antibodies against Bax, Bcl-2, procaspase-8, and Bid, indicating that apoptosis in HUVEC cells may involve a mitochondria pathway, by the demonstration of an increase in the Bax/Bcl-2 ratio. Together, our results suggest that the anti-angiogenesis activity of fascaplysin is through the direct effects of cell cycle arrest and apoptosis on HUVEC.