The Empirical Study Of Gene Therapy To Rabbit's VX₂ Hepatoma With Ad-P53 By Intervention Targeting Combined With Ultrasound-mediated Microbuble SonoVue

Objiective1. Making rabbit models of VX2 hepatoma in different hepatic lobes and to evaluate the effect on interventional intubation.2. Detecting the value of vascular endothelial growth factor(VEGF) in serum before and after infusing Ad-p53 gene, and the expression of p53 gene,matrix metalloproteinases 2(MMP2) and VEGF in carcinotisue. To investigate the effect of different interventional treatment on rabbit VX2 hepatoma.Methods1. A total of 20 rabbits was devided into 2 groups at random and 10 rabbits per group.20 rabbits were implanted VX2 tumor by two methods:inoculating in mid-right lobe and in mid-left lobe. Observing the growth conditions of VX2 tumor by ultrasound in the 7th,10th,14th day after inoculating, then to make angiography through hepatic artery.2. A total of 30 VX2 model rabbits was devided into 3 groups at random and 10 rabbits for each group. Micropipe was inserted nutrient artery of hepatoma through femoral artery before infusion. Physiological saline(control group),Ad-p53(group 2) were infused in defferent groups via nutrient artery of tumor and nutrient arteries were embolized with Lipiodol after infusion.The mixture of Ad-p53 and microbubble SonoVue(group 3) was infused via nutrient arteries of tumor with ultrasound radiation at equal pace, and with embolizing nutrient artery with Lipiodol after infusion. Samples of blood were acquired before and after infusion. VEGF was examined by ELISA and compared. Four days after infusion the rabbits were killed and the liver resected. The expressed protein of P53,MMP2 and VEGF was examined by immunohistochemistry(IHC).3. Statistical analysis:The statistical software:SPSS 11.0. The data was analyzed by Fisher's test and T test. The p value of less than 0.05 was considered to be statistical difference.Results1. The achievement ratio of different modeling wasn't different. Modeling in mid-right lobe of liver can ease interventional intubation.2. The results examined by ELISA show:the content of VEGF in serum after infusion was higher than that before infusion in group1,and lower in groups 2 and 3, and the decreasing scale of group3 is significant.3. Staining tumor tissues of groups1,2 and 3 by immunohistochemistry and measuring the value of OD of them, results show:The positive ratio of p53 in group 2 and 3 was higher vs group 1 respectively, and has significant difference(P<0.05). The positive ratio of VEGF and MMP2 in group 2 and 3 was lower vs group 1 respectively, and has significant difference(P<0.05) except for MMP2 in group 2 (P>0.05). The positive ratio of p53 in group 3 was higher than that in group 2(P<0.05), and the positive ratio of VEGF and MMP2 in group 3 was lower than that in group 2(P<0.05).Conclusions1. The method of modeling in mid-right lobe of liver can improve achievement ratio in interventional intubation.2. Infusing Ad-p53 gene via nutrient artery of tumor can improve expression level of p53, and supress expression level of VEGF and MMP2. It indicates indirectly that Ad-p53 gene can supress the growth of tumor. The method of infusing Ad-p53 combined with ultrasound-mediated microbubble was better than that of infusing Ad-p53 singly.