脱唾液酸糖蛋白受体介导的vp3基因靶向性治疗肝癌的实验研究

The Experimental Research on vp3 Gene Targeting Therapy for Hepa- toma by Asialoglycoprotein Receptor In Vivo

目的利用肝细胞表面存在特异的脱唾液酸糖蛋白受体(asialoglycoprotein receptor,ASGPR),探索ASGPR介导的vp3基因肝细胞靶向性治疗肝癌的方法。方法将携带vp3基因的质粒通过多聚左旋赖氨酸(poly-L-lysine,PLL)与该受体的天然配体脱唾液酸粘蛋白(asialoorosomucoid,Asor)结合,获得Asor-PLL-vp3复合物;通过体外转染、放射性同位素标记检测和动物实验鉴定该复合物的肝细胞靶向性。结果 成功制备了较纯的可溶性的蛋白-核酸复合物;体内外实验结果表明Asor-PLL-vp3复合物具有良好的肝细胞靶向性。结论通过制备Asor-PLL-vp3复合物,利用ASGPR介导实现了vp3的肝细胞靶向性基因转移,证实了该复合物具有体内靶向性治疗肝癌的可行性。

Objective By taking advantage of Asialoglycoprotein receptor ( ASGPR) existing on the surface of hepatocytes, an experiment was conducted on the vp3 gene hepatocyte targeting therapy for hepatoma mediated by ASGPR. Methods The recombinant DNA with vp3 gene was combined with asialoorosomucoid by poly-L-lysine (PLL) , and then the Asor-PLL-vp3 complex was obtained. The hepatocyte targeting of this complex was confirmed by gene transfection in vitro, radioactive isotope labelling and PCR results of genome DNA of different tissues of BALB/c mice. Results A pure soluble protein-DNA complex was successfully prepared. And the hepatocyte targeting of this complex was confirmed by those appraisements in vitro and in vivo. Conclusion The Asor-PLL-vp3 complex was successfully obtained and targeting gene transfer of vp3 gene mediated by ASGPR was achieved, thereby demonstrating the feasibility of targeting gene therapy for hepatoma by the complex.