Vasostatin Gene Therapy Suppresses The Angiogenesis Of Implant Hepatocellular Carcinoma In Nude Mice

Objective:Hepatocellular carcinoma is one of the most common malignances in China. Angiogenesis correlated with tumor biological behaviors have been affirmed. The characteristic of HCC which is abundant with blood supply and easy to metastasis and recurrence suggests that angiogenesis plays a crucial role in tumorigenesis. It has been found that vasostatin gene not only has a high expression on the surface of the noumenon tumor cells but also has a strong expression in the endotheliocytes of the new blood vessels in tumor tissue. In this study, we investigated the efficacy of vasostatin gene in treating subcutaneouly implanted hepatocellular carcinoma of nude mice and the effect to angiogenesis and apoptosis with immunohistochemical staining. This research offers us a novel idea of gene therapy in HCC.Methods:1. vasostatin/pcDNA3.1 were transformed into competent E. coli cells and assayed after endonuclease cutting.2.60 male nude mice which were 4 weeks old were separated into 2 groups in random and each group had 15. HCC cell line HepG2 was transplanted subcutaneous in nude mice, then the vasostatin gene were transfected into the tumor with electroporation. Then we observed the growth of the tumors. After 6 weeks, we took the tumors, weighted them and calculated the tumor inhibit rate.3.Protein vasostatin gene and CD31 were examined with the immunohistochemical staining respectively. The expression of vasostatin gene and tumor microvessel density (MVD) were assessed.4.TUNEL (TdT-mediated biotinylated-dUTP nick end labling method) was adopted to analysis apoptosis of tumor cells and the AI (apoptosis index) was calculated. Further, the AI of every group was compared.Results:1. The bulks of tumors were obviously different among each group. Tumors in the control group were bigger than those in the other groups. Tumors in the vasostatin group were the smallest. The tumor weight of each group was significantly different (P<0.05).2.The MVD of the control, vasostatin group was 18.26±1.72,5.62±0.59. It was obviously that the MVD of vasostatin group was lower than that of the control group (P<0.05).3.The AI of the control, vasostatin group were 6.25±1.32,16.48±0.96 respectively and had significant difference among them(P<0.05).Conclusions:1.The antisense gene of vasostatin can markedly inhibit the angiogenesis of tumor.2.The antisense gene of vasostatin can induce cell apoptosis significantly. 3. The antisense gene of vasostatin can suppress tumor growth.