Inhibitory Effect Of Bd0801 Or BD0801 Combined With 5-FU On The Proliferation Of HCC Cell Line SMMC-7721and Angiogenese In Vitro

Objective:To investigate the inhibitory effect and mechanism of BD0801 single or combined with 5-fluorouracil(5-Fu) on the human hepatocellular cell line SMMC-7721 proliferation and anti-angiogenesis effect in vitro.Methods:In vitro, choose the human hepatocellular cell line SMMC-7721 and the human umbilical vein cell line EAhy926 as experimental object, with Bevacizumab as positive agent, analyze the cell proliferation in different concentration of SMMC-7721 and EAhy926 by MTT. Make use of the flow cytometry to detect the changes of apoptotic rate and cell-cycle. Analyses of VEGF content in cell media, and VEGF and pVEGFR expressions were obversed by enzyme-linked immuno sorbent assay(ELISA) and Western blot, respectively. Use the wound healing and Transwell to investigate the inhibitory effect on the migration of EAhy926. Test the influcence on the angiogenesis of EAhy926 with the tube formation assay.Results:1. BD0801 inhibit the growth of SMMC-7721 and EAhy926 cells, induce the cells to apoptosis, and arrest the cell cycle at G0/G1 phase, In vitro. Meanwhile, BD0801 down-regulate the level of VEGF in cell culture medium, inhibit the abilities of EAhy926 cells’with migration(wound healing and transwell assays) and assembly into vascular structures (tube formation assay).2. Compare with the single agents,it is the synergistic interaction between BD0801 with 5-Fu, and it is more effective than the combination with Bev and 5-Fu, futher increased the cellular proliferation inhibition rate, apoptosis rate and arrested the cell cycle of SMMC-7721 and EAhy926, inhibition rate of EAhy926 cells migrate and tube. There was a stronger decrease in VEGF release and the protein.Conclusions:First, BD0801 targeting VEGF/VEGFR is able to inhibit cell proliferation, induce apoptosis and arrest cell cycle on SMMC-7721 and EAhy926 cells, as well as seriously effect angiogenesis of EAhy926 in vitro. Second, BD0801 in combination with 5-Fu can produce synergistic effect on SMMC-7721 and EAhy926, including inhibiting proliferation, inducing apoptsis, regulating cell cycle as well as inhibiting VEGF/VEGFR pathway on the two cell lines and angiogenesis of EAhy926. Third, BD0801 combination with 5-Fu has stronger inhibitory effect than Bev combination with 5-Fu. It is showed that BD0801 might be a potential and promising agent of anti-VEGF monoclonal antibody to be further study in HCC therapy.