论文部分内容阅读
如何使得外源基因稳定地从吞噬溶酶体中释放 ,是提高受体介导基因转移系统转移效率的关键。本文以绿色荧光蛋白质粒为报告基因 ,合成针对表皮生长因子受体 (EGFR)的相应 16肽配体寡肽和流感病毒血凝素HA2 0寡肽 ,并与多聚赖氨酸连接 ,连接物与绿色荧光蛋白报告基因按 1∶1混合 ,构建新型肝癌靶向性转移系统 ,命名为四元复合体。分别以四元复合体和脂质体在体内外进行基因转染 ,流式细胞仪和激光共聚焦显微镜检测绿色荧光蛋白表达。结果表明 ,四元复合体介导基因定向转染至肿瘤细胞 ,其转染率为 44 95 % ,显著高于脂质体转染组( 3 3 0 9% ,P <0 0 5 ) ,动物实验证明 ,四元复合体介导绿色荧光蛋白特异性表达于肿瘤细胞 ,具有良好的应用前景
How to make the exogenous gene stably released from phagolysosomes is the key to improve the efficiency of receptor mediated gene transfer system. In this study, green fluorescent protein plasmid was used as a reporter gene to synthesize the corresponding 16 peptide ligand oligopeptide against epidermal growth factor receptor (EGFR) and the influenza virus hemagglutinin HA20 oligopeptide and to link with polylysine. And green fluorescent protein reporter 1: 1 mixed to build a new liver cancer targeting metastatic system, named quaternary complex. Gene transfection was performed in vitro and in vivo with quaternary complex and liposome respectively. The expression of green fluorescent protein (GFP) was detected by flow cytometry and laser confocal microscopy. The results showed that the transfection rate of the quaternary complex-mediated gene transfection to tumor cells was 44 95%, which was significantly higher than that in the liposome-transfected group (33.09%, P <0.05) Experiments show that the quaternary complex-mediated green fluorescent protein-specific expression in tumor cells, has a good prospect