Experimental Study Of Inhibitory Effect Of Polypeptide Extract Scorpion Venom On Angiogenesis Of HepG2Heptocecullar Carcinoma

ObjectiveTo study the effects of polypeptide extract scorpion venom (PESV) on angiogenesisassociated factors in tumor micro-environment.To observe the inhibitive effects of PESVon tumor growth.To study the anti-angiogenic molecular mechanism,and further reveal theanti-tumor mechanisms of PESV.Provide the experimental basis for the treatment ofhepatocellular carcinoma.Therefore PESV can be combined with traditional chemotherapydrugs.Methods1.MTT assay was used to test the inhibition of cell proliferation．Flow cytometry wasused to detect cell cycle.2.Hepotocecullar carcinoma models was established by inoculating with HepG2cellssuspension in right armpit subcutaneously.The tumor-bearing mice were randomly dividedinto three groups:the tumor-bearing group,PESV group and the positive control group.Thetumor volume and tumor inhibitory rate were determined.The inhibition effect of PESV ontumor growth was observed.3.Conventional HE was done,and tumor tissue pathological changes were observedwith light microscopy.4.The expression of VEGF,HPA,survivin and CD34in tumors were determined byimmunohistochemistry-staining.5.The protein expression of VEGF,HPA and survivin in tumors were detected bywestern-blot. Results1.The effect of PESV on the inhibition of cell proliferation and cell cycleCompared with the negative control group,when PESV＜20μg mL-1,the inhibitiveeffect on tumor cells was not obvious.But when PESV＞40μg mL-1,the proliferationinhibition increased significantly. Flow cytometry results showed that, PESV enabled cellcycle arrest at G0∕G1phase, to reduce the proportion of cells in S and G2∕M phase,compared with the negative control group,the difference was significant.2.Effects of PESV on tumor growth of HepG2hepatocecullar carcinomaIn the PESV group and the positive control group,the tumor weights of mice weresignificantly reduced,and the inhibition effect of PESV on tumor volume was obvious(P＜0.05).3.Effects of PESV on expression of VEGF,HPA,survivin and CD34in tumor tissueThe results of immunohitochemistry-staining showed that,positive products ofVEGF,HPA and survivin were mainly localized in the cytoplasm of tumor cells.By thesemi-quantitive image analysis.The expressions of VEGF,HPA and survivin in PESV groupand the positive group were significantly lower than that in the control group(P＜0.05).4.Effects of PESV on VEGF、HPA and survivin protein level in tumor tissueThe VEGF、HPA and survivin protein expression can be detected in tumor-bearinggroup,PESV group and the positive group.The protein expression of VEGF、HPA andsurvivin in PESV group and the positive group were significantly lower than that intumor-bearing group.The differences were statistically significant(P＜0.05).Conclusions1.PESVcan inhibit the growth of hepotocecullar carcinoma.2.PESV has an inhibitive effect on the growth of hepatocecullar carcinoma.3.PESV can intervene angiogenesis of tumors,the mechanism may be associated withreducing the expression of VEGF,HPA and survivin and the micro-vessel desity.