Experimental Study Of The Anti-Tumor And Anti-Angiogenesis Of Cinobufacini And Combined With AS₂O₃

Objective: By experimental study, we explored the effects and mechanisms of anti-tumor and anti-angiogenesis of cinobufacini combined with AS₂O₃.Methods:In vitro studies, human hepatocellular carcinoma cell line SMMC-7721 and human umbilical vein cell line ECV-304 were treated with cinobufacini combined with AS₂O₃, then their growth was analyzed by MTT assay and transmission election microscopy dectection. Using immunohistochemical staining, the change of VEGF,EGFR`s expression in SMMC-7721 and ECV-304 were investigated. By the method chick chorioallantoic membrane (CAM) technology, we investigated the inhibitory effect of cinobufacini combined with AS<sub>2</sub>O<sub>3</sub> on tumor neovascular. In vivo, using the nude mice with the high metastatic human HCC transplanted; 32 nude mice was divided into 4 groups(NS group, cinobufacini group, AS<sub>2</sub>O<sub>3</sub> group, cinobufacini combined with AS<sub>2</sub>O<sub>3</sub> group), 8 mice in each group. Experiment started on the 7th day after the tumor transplantation. All groups were treated with abdominal injection per day for 3 weeks, the killed and dissected, drug`s inhibitory effect on the transplantation tumor was observed, and the changes of VEGF,EGFR and MVD were investigated by using immunohistochemical staining.Results: In vitro studies showed, cinobufacini could inhibit the growth of human hepatocellular carcinoma cell line SMMC-7721 and human umbilical vein cell line ECV-304. The inhibited effect to SMMC-7721 is enhanced by cinobufacini combined with AS₂O₃, but the inhibited effect to ECV-304 is not enhanced by cinobufacini combined with AS₂O₃. Cinobufacini down-regulated VEGF and EGFR`s expression of SMMC-7721 and ECV-304 and the effects are enhanced by cinobufacini combined with AS<sub>2</sub>O<sub>3</sub>. Cinobufacini could inhibit the neovascular in the CAM and the effect are enhanced by cinobufacini combined with AS<sub>2</sub>O<sub>3</sub>. In vivo studies showed cinobufacini inhibited the transplanted tumor significantly and this effects are enhanced by cinobufacini combined with AS<sub>2</sub>O<sub>3</sub>. Cinobufacini could drow-regulated VEGF and EGFR`s expression and decreased the MVD, and the effects are enhanced by cinobufacini combined with AS₂O₃.Conclutions: Cinobufacini can inhibite the HCC and has anti-angiogenesis effect. The combination of cinobufacini and AS₂O₃ might produce additive effect. The mechanism of anti-angiogenesis probably is that cinobufacini can inhibite the growth of endothelial cell and down-regulate the expression of VEGF,EGFR.