Study Of Antitumor Effect And Mechanism Of RAdinbitor

Objective:To study the anti-tumor effect and mechanism of rAdinbitor, rooted in a novel disintegrin from the snake venom of Agkistrodon halys brevicaudus stejneger, so as to provide necessary experiment and theory evidences for its clinical experiment in the future.Methods:The E.coli BL21 contained the rAdinbitor gene previously construted by the use of genetic engineering technology was cultured generously. After broke by ultrasound, total protein in the bacteria was extracted and the rAdinbitor was purified by His-BindColumn affinity chromatography. The protein was identified with 18% Tricine-SDS-PAGE.Twelvty of BALB/C mice were randomly divided into two groups, oral administration and intramuscular administration group, a half female and a half male in each group. The animals of each group were divided into five subgroups, one-second female and one-second male in two groups.1×106 of H22 hepatoma cells were inoculated into the right axillary hypoderm of each experimental BALB/C mouse to establish the required tumor animal model. The treatment dose of rAdinbitor was 0.5mg/kg, 1.25mg/kg and 5mg/kg weight respectively. Physiological saline was used as negative control agent (0.2ml/kg weight) and the recombinant Human Endostatin as positive control agent (1.65mg/kg weight). Tumor tissues were taken out from the killed experimental mice on the twenty-first day and each tumor tissue was weighed. Tumor tissues slices stained with HE were observed and counted the number of blood vessel under microscope.Results:The rAdinbitor was over-expressed in E.coli BL21. A great quantity of purified rAdinbitor protein was successfully obtained by affinity chromatography. The animal experiment showed, compared to the negative group, that the weight and the number of blood vessel of tumor tissue in each experimental group was reduced significantly (P<0.05).Conclusion:The rAdinbitor, rooted in a novel disintegrin from the snake venom of Agkistrodon halys brevicaudus stejneger, could remarkably inhibit the growth of H22 hepatoma cells. It could be speculated that the mechanism of rAdinbitor's inhibitory effect on tumor growth is that it may reduce the angiogenesis around tumor tissue.