Experimental Study Of The Combined Effect Of Adenovirus Containing CD/TK Fusion Gene Driven By VEGF Promoter Mixed With Lipiodol Targeted On VX2 Hepatocarcinoma Of Rabbits

PurposeTo evaluate the combined therapeutic effect of recombinant adenovirus CD/TK fusion gene driven by vascular endothelial growth factor prompter system(Ad-VEGFp-CDglyTK) mixed with ultra-fluid lipiodol, which is injected via hepatic artery,targeted on transplantation VX2 hepatocarcinoma rabbits. Materials and MehtodsPART ONE To construct the recombinant adenovirus CD/TK fusion gene driven by vascular endothelial growth factor prompter system (Ad-VEGFp -CDglyTK).PART TWO Establishment of the Transplantation VX2 Tumor Model of hepatocarcinoma in rabbits.PART THREE VX2 carcinoma cells were surnically implanted into the left liver lobe in 40 male New Zealand white rabbits, which were randomly divided into 4 groups with 10 rabbits each.Normal saline(control group);ultra-fluid lipiodol(LP group); Ad-VEGFp-CDglyTK system,then intraperitoneal treatment with GCV and 5-FC(Ad-VEGFp-CDglyTK group) and Ad-VEGFp-CDglyTK system mixed with ultra-fluid lipiodol, then intraperitoneal treatment with GCV and 5-FC(LP+ Ad-VEGFp-CDglyTK group)were respectively infused via hepatic catheter for each group.The tumors' sizes were examined before and 10,15 days after the procedure by MRI,SCT, DSA or US. With the RT-PCR, we examined the expression of sucide genes on the mRNA level.The survival period and serum CBC,ALT,γ—GT, Cr, chol, TG level were observed. Anti-Human VEGF and Anti-Human FactorⅧRelated Antigen expression were estimated by immunohistochemistry. The microvessel density(MVD) were caculated. Hepatic apoptosis was assessed by In Situ Cell Death Detection Kit, POD(Roche) staining and the TUNEL method.Statistical analyses: All data were expressed as (?)±S. Differences between groups were assessed with SNK-q test. All statistics were computed by using SPSS11.5 statistical package. P＜0.05 was considered to indicate a statistically significant difference.ResultsThe suicide genes were all expressed in the tumors.The tumor growth rate in LP+ Ad-VEGFp-CDglyTK group was lower than that in LP, Ad-VEGFp-CDglyTK group and control groups(P＜0.05).The survival period of LP+ Ad-VEGFp-CDglyTK group was 53.0±3.0d, which was significantly higher than that of other two study nroups(P＜0.05). The change of serum AST level after the procedure showed no significant difference between LP+ Ad-VEGFp-CDglyTK group and other two study groups(P＞0.05), although it was significantly higher than that of control nroup(P＜0.05). VEGF mRNA in cancerous tissues was shown most in LP group(P＜0.05). There was no significant difference among other three groups. The MVD in LP+ Ad-VEGFp-CDglyTK group was significantly less than that of other groups. Compared with the group without Ad-VEGFp-CDglyTK, the percentage of apoptotic hepatocytes after CC14 intoxication was significantly lower, as was ALT activity. In addition, ALT activity normalised more rapidly in the LP+ Ad-VEGFp-CDglyTK group.ConclusionsThe use of Ad-VEGFp-CDglyTK system mixed with ultra-fluid lipiodol as interventional chemoembolization agent can greatly decrease tumor growth rate and prolong the survival period, inhibit Anti-Human VEGF and Anti-Human Factor VIII Related Antigen expression and microvessel density.Moreover, the related hepatic damage is reversible.