Experimental Study On Anti-angiogenic Effect In Hepatocellular Carcinoma Of Recombinant Human Endostatin Combined With Oxaliplatin Or Cinobufacini

Objective:Observe the effect to suppress the growth of H22liver tumor model and safety of recombinant human endostatin (Endostar) work together with Oxaliplatin or Cinobufacini, and investigate the mechanism.Methods:Establish mouse subcutaneous H22liver tumor model. It is about one week for the tumor reach to100mm3.Part1:ninty-six mice with subcutaneous H22liver tumor were divided into six groups randomly as follows:Control group (Normal saline); Oxaliplatin (dl-3) group; Endostar (d1-7) group; Endostar+Oxaliplatin (dl-3) group; Endostar+Oxaliplatin (d4-6) group; Endostar+Oxaliplatin (d7-9) group. Normal saline and Endostar were administrated subcutaneously. Oxaliplatin was administrated intraperitoneally.(1) Recorded the volume of tumor.(2) Measured VEGF of the concentration of serum and expression of MVD in transplanted tumor.(3) Measured the Even blue content of the tumors with the same weight, which can reflect chemotherapy content of the tumor.(4) Recorded the body weight of the mice. Test the alanine transaminase and aspartate transaminase in the blood, Observed the pathological changes of liver and kidney under the microscope.Part2:Forty mice with subcutaneous H22liver tumor were divided into four groups randomly as follows:Control group (Normal saline); Cinobufacini group; Endostar group; Endostar+Cinobufacini group. The Endostar and Normal saline were administrated subcutaneously for14days. Cinobufacini was administrated intraperitoneally for14days.(1) Recorded the volume of tumor.(2) Measured VEGF of the concentration of serum and expression of MVD in transplanted tumor.(3) Recorded the body weight of the mice. Test the alanine transaminase and aspartate transaminase in the blood, Observed the pathological changes of liver and kidney under the microscope.Results:Part1:(1) The volume of the transplanted tumor of Endostar+Oxaliplatin (dl-3) group and Endostar+Oxaliplatin (d4-6) group are significantly lower than that of the control group. The others could not be controlled effectively.(2) Compared with the control group, Endostar+Oxaliplatin (dl-3) group, Endostar+Oxaliplatin (d4-6) group and Endostar+Oxaliplatin (d7-9) group lowered VEGF protein level and the expression of MVD significantly.(3) On the4th day, the Even blue content of the transplanted tumor of Endostar group was a bit higher than the control group. On the7th day, the Even blue content of the transplanted tumor of Endostar group was lower than control group. On the10th day, The Even blue content was nearly the same in both groups.(4) The weights of the mice of each group are almost the same. The ALT and AST test, pathological changes of liver and kidney were normal.Part2:(1)The volume of the transplanted tumor of Endostar+Cinobufacini group are significantly lower than that of the control group. The others could not be controlled effectively.(2) Compared with the control group, Cinobufacini group and Endostar+Cinobufacini group can lowered VEGF protein level significantly. Combination group can lowered the expression of MVD significantly.(3)The weights of the mice of each group are almost the same. The ALT and AST test, pathological changes of liver and kidney were normal.Conclusion:Part1:(1) Using Oxaliplatin followed by using Endostar in the first3days and using them in the first time both can suppress the growth of transplanted tumor, and the fomer one is better.(2) Endostar could inhibit the level of VEGF protein and MVD effectively combined with Oxaliplatin. But it can’t inhibited the level of VEGF protein and MVD effectively by itself.(3) On the4th day, the Even blue content of the transplanted tumor of group Endostar was a bit higher than control group’s. On the7th day, the Even blue content of the transplanted tumor of Endostar was lower than control group. On the10th day, the Even blue contents were nearly the same in both groups. The potential mechanism may be Endostar has normalized vasculature of H22liver tumor on the4th day.(4) The usage of Endostar and Oxaliplatin is safety for mice to cure subcutaneous H22liver tumor.Part2:(1) Using Endostar or Cinobufacini respectively cannot suppress the tumor effectively. However, using them together can work observably.(2) Cinobufacini can suppress the expression of VEGF. Endostar and Cinobufacini not only can suppress the expression of VEGF observably, but also can decrease the expression of VEGF and MVD.(3) The usage of Endostar and Cinobufacini is safety for mice to cure subcutaneous H22liver tumor.