摘要
目的研究转铁蛋白(Tf)对p53基因转染效率的影响,并探讨介入技术与Tf联合应用对基因治疗肝癌的双重靶向作用。方法应用pCMV-myc-p53质粒与阳离子脂质体LipofectAMINE复合物,转染3种肝癌细胞株LM6、Hep3B、YY以及正常肝细胞株L02,以不同浓度Tf(0、10、25、50及100μg)介导转染,蛋白印迹法检测各细胞株中p53表达情况,并对Tf影响p53转染效率的关系进行分析。再建立兔VX2肝癌动物模型,经介入导管输注Tf-质粒-脂质体复合物,提取肿瘤组织蛋白,蛋白印迹法检测P53表达。结果4种细胞应用p53-脂质体复合物以及不同浓度Tf转染后48h的蛋白印迹法检测,发现Tf在10～100μg可明显增强p53-脂质体的转染效率,且转染效率随Tf浓度增加而增强。动物模型组织提取蛋白检测结果,显示Tf明显增强P53蛋白表达。结论Tf能增强脂质体-基因转染,介入技术与Tf联合双重靶向对肝癌的基因治疗具有相当的应用前景。
Objective To investigate the function of transferrin-DNA complex, transported by transferrin （Tf） and trans-arterial injection via interventional approach be the duel-target-orientated delivery and the transferring into malignant cells to get more effective therapy. Methods p53-LipofectAMINE ligand with different concentrations of Tf （0, 10, 25, 50, 100 μg） transfected the 4 strains including LM6、Hep3B、YY and L02 in vitro to evaluate the gene transfection efficiency through western blot. Then, after setting up the VX2 hepatocarcinoma models, we delivered the Tf-p53-LipofectAMINE complex into the hepatic arteries via interventional techniques to analyse the transfection efficiency in vivo. Results Tf, within the range of 10～100 μg, could increase gene transfection efficiency mediated by liposome, and the efficiency increases with the raise of Tf concentration. Combination with interventional technique to inject Tf-DNA complex into tumor arteries, gene transfection efficiency was enhanced in rabbit models. Conclusion Tf can enhance gene liposome transfection efficiency, furthermore with combination of interventional catheter technique, there would be a potential duel-target-orientated gene therapy method.
出处
《介入放射学杂志》
CSCD
2007年第2期99-103,共5页
Journal of Interventional Radiology
