Inhibitory Effect Of BD0801 Or BD0801 Combined With 5-FU On The Proliferation Of HCC Cell Line HepG2 And Human Umbilical Vein Endothelial Cell Line In Vitro And Its Mechanisms

Objective:To investigate the influence of humanized anti-VEGF monoclonal antibody BD0801or BD0801combined with 5-fluorouracil(5-FU) on the proliferation inhibition effect of human hepatocarcinoma cell line HepG2 and human umbilical vein endothelial cell line (HUVEC), and it's related mechanisms.Methods:In vitro, inhibitory effect, interaction effect as well as the related mechanism of human hepatocarcinoma cell line HepG2 and HUVEC induced by BD0801or BD0801 combined with 5-FU at various concentration for 0-72h were observed by 3-(4,5-dimethylthiazole-2-yl)-2, 5-diphenyl tetrazolium bromide(MTT) assay,4,6-Diamidino-2-phenylindole (DAPI) staining, flow cytometry assay, enzyme-linked immunosorbent assay(ELISA),and Western-blot.Results:In vitro, compared with Bevacizumab (Bev), BD0801 could effectively inhibit the growth of human hepatocarcinoma cell line HepG2 and HUVEC. After incubated with BD0801 the cells showed typical morphological apoptosis and biochemical characteristic such as chromosome condensation, chromosome margination and nuclear fragmentation. Meanwhile, significant apoptosis-inducing effect was found in FCM, the cell cycle was arrested at G0/G1 phase, the level of VEGF in the cell culture medium was down-regulated. Compared with BD0801, 5-FU,or Bev combined with 5-FU drug groups, BD0801 combined with 5-FU obviously increased the inhibitory rate of human hepatocarcinoma cell line HepG2 and HUVEC. There was a significant difference between combined medication group and monotherapy group. The effect of inducing apoptosis can be increased and the proportionality of cells at S phase was increased following the treatment with BD0801 combined with 5-FU. It is showed that the level of VEGF in the cell culture medium was down-regulated and VEGF/VEGFR signal path was inhibited obviously.Conclusions:1. BD0801 has significant inhibitory effect of proliferation on human hepatocarcinoma cell line HepG2 and HUVEC in vitro, which may be related to inducing the cell apoptosis, regulating cell cycle as well as inhibiting VEGF/VEGFR signal path.2. BD0801 combined with 5-FU may obviously improve the inhibitory effect of proliferation on human hepatocarcinoma cell line HepG2 and HUVEC in vitro, and the interaction is synergistic, whose main mechanism is to increase apoptosis-inducing effect, further regulating cell cycle, and inhibit VEGF/VEGFR signal path.