表皮生长因子受体介导的新型肝癌靶向性基因转移

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

EGFR-mediated gene delivery targeting to hepatocarcinoma cells

To construct a gene delivery system targeting hepatocarcinoma cells via EGFR.GE7,a 16-peptide molecule specific to EGFR, and HA20,a homologue of N-terminus of haemagglutinin of Influenza Viral envelope protein were synthesized and conjugated with polylysin. The conjugate was mixed to report plasimd pEGFP-N1, a GFP expression vector, to construct a novel gene delivey system named as 4-element complex. In vitro and in vivo gene transfection were respectively made by 4-element complex and lipofectin. Laser confocusial microscopy showed that 4-element complex mediated GFP expressed in Hepatocarcinoma cell line HepG2.2.15 but not in human umbilical vein endothelial cell line ECV304. Flew cytometry assay showed that the transfection rate of 4-element was 44.95% that was greater than that of lipofectin(33.09%,P<0.05). BalB/c nude mice bearing HepG2.2.15 cells were injected with 4-element complex and lipofectin containing report gene via trail vein when the tumor size was about 0.5cm in diameter. After 3 days, specimens were collected and laser confocusial microscopy indicated that GFP only expressed in hepatoma cells of nude mice treated with 4-element complex. However, GFP expressed in all tissue sampes of lipofectin treated nude mice.