经肝动脉注射双腺病毒载体诱导Bax基因在肝脏靶向表达实验探讨

Experimental study of Human Hepatocellular Carcinoma with Bax Gene Therapy via Hepatic Artery Delivery

目的:探讨Bax基因对肝癌细胞促凋亡作用及转基因途径对转染效率的影响。方法:双腺病毒载体系统介导Bax基因转染人肝癌细胞系QGY7703,并将Bax基因通过鼠尾静脉、肝动脉进行转染,观察细胞凋亡及不同转染途径对靶器官的影响。结果:1)Bax基因转染导致人肝癌细胞凋亡。实验组凋亡百分率明显高于各对照组。2)肝动脉插管组LacZ在靶肝细胞转染率高;非靶器官分布表达少,靶/非靶比值大(P<0.05);阻断血供有利于以较小病毒滴度获得较高的转染率;尾静脉组靶肝细胞转染率低,在非靶器官分布表达多。结论:应用肝动脉超选择技术,经肝动脉传输Bax基因治疗肝癌具有安全、低毒和靶向选择性,为Bax经肝动脉基因治疗提供了实验依据。

Objective: To investigate the hepatocellular carcinoma cell apoptosis induced by Bax gene and the effect of different ways on genes delivery. Methods: The anti-hepatocellular carcinoma activity of Bax gene transferred to the human hepatocellular carcinoma cell line QGY7703 was observed. Bax gene was transferred via murine caudal vein or hepatic artery to investigate the different effects on target organ and non-target organ. Results: 1) The Bax gene mediated by biadenoviral vector could induce apoptosis in human hepatic carcinoma cell QFY7703. The cell apoptotic rate in experimental group(Bax) was 50.2±6.9% while 32.1±9.7% in Ad/CMV-p53 group, which showed that Bax apoptotic rate was significantly higher than that of p53 and other control group. 2) The gene distribution and transduction rate in superselective group was higher ( the max rate 70%, average rate 36.33± 6.11%) than that in control group (P<0.05). In addition, the gene expression in non-target organ was fewer and target/non-target rate was high; high transduction can be accomplished with low virus titer through hepatic artery block; compared to hepatic artery, the hepatic cell transduction rate was lower when delivered through caudal vein. More gene distributed to non target organs. Conclusions: The superselective hepatic artery delivery with Bax gene therapy is safe, low toxic, specific and effective. This study provides the basis for Bax gene therapy via hepatic artery.