title: Direct effects of fascaplysin on human umbilical vein endothelial cells attributing the anti-angiogenesis activity.
abstract: 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 2.6 μM 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 36 h, comparing to the 22.82 ± 1.2% in methanol treated cells. Apoptosis of HUVEC cells was induced by 1.3 μM 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.
authors.affiliation: Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, P R China.
authors.forename: Y L
authors.initials: YL
authors.lastname: Zheng
authors.forename: X L
authors.initials: XL
authors.lastname: Lu
authors.forename: J
authors.initials: J
authors.lastname: Lin
authors.forename: H M
authors.initials: HM
authors.lastname: Chen
authors.forename: X J
authors.initials: XJ
authors.lastname: Yan
authors.forename: F
authors.initials: F
authors.lastname: Wang
authors.forename: W F
authors.initials: WF
authors.lastname: Xu
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D053148:Caspase 3 / Q000378:metabolism; D018929:Cell Culture Techniques; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D005434:Flow Cytometry; D016193:G1 Phase / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D020287:In Situ Nick-End Labeling; D007211:Indoles / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D013997:Time Factors; D014471:Umbilical Veins / Q000166:cytology; D051028:bcl-2-Associated X Protein / Q000378:metabolism; D000644:Quaternary Ammonium Compounds; D002243:Carbolines; D007212:Indolizines
chemical_list: D020533:Angiogenesis Inhibitors; D053148:Caspase 3; D007211:Indoles; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; C494826:BAX protein, human; C069346:fascaplysine; D000644:Quaternary Ammonium Compounds; D002243:Carbolines; D007212:Indolizines