| Objective: Primary hepatic carcinoma is one of the most common malignances in China. The majority of pathologic type of hepatic carcinoma is hepatocellular carcinoma (HCC) which is beyond 90%. We have examined angiogenesis correlated with tumor biological behaviors. The characteristic of hepatic carcinoma 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 integrin αvβ3 not only has a high expression on the surface of the solid tumor cells but also has a strong expression in the endotheliocytes of the new blood vessels in the tumor tissue. For these reasons, it has become a focal point in recent years that cross-links corresponding radionuclide or chemical medicine to the monoclonal antibody or antagonist peptide against integrin αvβ3 to carry out targeting diagnosis or treatment on the tumor. In this study, our aim is to explore the efficacy of antisense integrin avp3 in treating implant hepatocellular carcinoma of nude mice and the effect to angiogenesis and apoptosis.Methods: AlphaV/pcDNA3 and Beta3/pcDNA3 were transformed into competent E. coli cells and assayed after endonuclease cutting. 120 male nude mice which were 4-6 weeks old were separated into 4 groups in random and each group had 30. HCC cell line HepG2 was transplanted subcutaneously in nude mice, the mixture of antisense integrin av,p3 and Fugene6 was transfected into the tumor. Then we observe the growth of the tumors. After 6weeks, we take the tumors, weight them and calculate the tumor inhibit rate. av,p3 protein and CD31,VEGF were examined by immunohistochemistry respectively. Tumor microvessel, density (MVD) was assessed using the expression of CD31. TUNEL ( TdT-mediated biotinylated-dUTP nick end labling method) was adopted to analysis apoptosis of tumor cells.Results: (1) The volumes of tumors were obviously different among each group. Tumors in the contrast group were bigger than those in the other groups. Tumors in the av(33 group were the smallest. The difference may be found easily through the growth curve. The tumor weight of each group was significantly different(P<0.05), and that ofavp3 group was lighter than those of av,P3 group (P<0.05). The tumor inhibit rate of av group,(33 group and av^3 group was 6.87%,5.71%,21.41% respectively.(2) Protein of integrin av and (33 expressed in all groups. In the contrast group, 20 cases were positive and the positive rate was 66.67%. The positive rate of av group,p3 group and avp3 group was 30.00%, 26.67%, 26.67% respectively and no difference was found among them(P>0.01), but significantly lower than that of the contrast group(P<0.001). ( a '=0.05/6=0.0083).(3)The MVD of the contrast group, av group, P3 group and avP3 group was 17.53+1.88,16.06+1.92,15.83+2.00,14.86+1.69. It was obviously that the MVD of av group, P3 group and avp3 group was evidently lower than that of the contrast group(P<0.01), and the difference between avp3 group and av group, p3 group was found among them(P<0.05).(4) All the cases expressed VEGF. In the contrast group, the positive rate which had significant difference with av group, p3 group and avp3 group(P<0.005)was 60.00%. The positive rates of av group, p3 group and avp3 group was 20.00%> 23.33%, 16.67% respectively and had no difference among themCi^O.OlM a '=0.05/6=0.0083).(5) The AI of the contrast group, av group, p3 group and avP3 group waslO.533+3.29%, 19.80+4.06%, 21.93+3.26%, 24.03+4.45% respectively andhad significant difference among them(P<0.01). The AI of av03 group was higher than that of av group and 03 group (P<0.05), meanwhile, the AI of 03 group was higher than that of av group(P<0.05).Conclusions: (1) The antisense gene of integrin av03 can apparently inhibit the growth of tumor.(2) The antisense gene of integrin av|33 may inhibit the angiogenesis of tumor.(3) The antisense gene of integrin av03 could accelerate cell apoptosis.(4) It is more efficient when the antisense gene of integrin av with the antisense gene of integrin 03 are combinely used.
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