转铁蛋白-多聚乙烯亚胺体外转染小鼠骨肉瘤细胞的实验研究

Transferrin-polyethyelenimine as a tumor-targeting vector for gene transfer to murine osteosarcoma

目的评价转铁蛋白-多聚乙烯亚胺(transfertin-polyethylenimine,Tf-PEI)靶向性转染小鼠骨肉瘤细胞的可行性,并确定最佳的转染条件。方法以Tf-PEI为载体,将绿色荧光蛋白基因和荧光素酶基因分别导入小鼠骨肉瘤细胞,用荧光显微镜及荧光素酶检测系统检测其转染效率。以游离转铁蛋白竞争性拮抗Tf-PEI,并观察它对转染效率的影响。结果Tf-PEI可以高效的、靶向性的转染小鼠骨肉瘤细胞。当N/P比值为5时,Tf-PEI的转染效率最高。游离转铁蛋白能够显著的拮抗Tf-PEI的转染。结论Tf-PEI是一种高效率、低毒性、肿瘤靶向性的基因转染载体,它可以作为体外恶性肿瘤基因治疗试验的一个有效工具。

Objective To investigate the feasibility and efficiency of transferrin- polyethylenimine （Tf-PEI） for gene transfer to murine osteosarcoma. Methods Using Tf-PEI as vector, murine osteosarcoma cells were transfected with plasmid DNA encoding for green fluorescent protein （GFP） and luciferase. Expression of the transduced gene was monitored by fluorescence microscope and Iuciferase assay system, Using free transferrin to inhibit Tf-PEI, the influence to the transfection of Tf-PEI was observed, Results Tf-PEI can effectively and specifically deliver gene into murine osteosarcoma. When N/P ratio equaled to 5, the transfection efficiency of Tf-PEI was the highest. The transfection can be remarkably inhibited by free transferrin. Conclusion Tf-PEI is a high efficiency and low cytotoxicity tumor-targeting vector. It can be used in the area of tumor-targeting gene therapy in vitro as an effective tool,