脂质体介导法转染肿瘤细胞效率的优化  

Optimization of Transfection Efficiency of Tumor Cell Mediated by Liposome

作　　者：潘宁[1] 章蔼然[1] 侯颖春[1] 

机构地区：[1]陕西师范大学生命科学学院肿瘤细胞分子生物学实验室,陕西西安710062

出　　处：《生物技术》2008年第6期47-50,共4页Biotechnology

基　　金：西安市科技局社会发展研发项目资助("AnnexinⅡ基因敲除Caco-2细胞系的建立和初步应用";YF07188)

摘　　要：目的：研究优化影响脂质体转染效率的因素，以提高脂质体转染效率，为相关研究和应用提供参考。方法：以绿色荧光蛋白（GFP）作为报告基因，采用脂质体Lipofectamine2000包裹pU6H1-GFP-FAK重组质粒转染Caco-2细胞，研究了细胞接种密度、DNA用量、脂质体与DNA的比例、脂质体-DNA复合物的形成时间、细胞与脂质体复合物的孵育时间、血清的有元及细胞的传代次数等因素对脂质体转染效率的影响。结果：2—5次细胞传代，2×10^5接种密度、4μgDNA用量、2．5：1的脂质体与DNA比例、30min脂质体-DNA复合物形成时间以及6h细胞与复合物孵育时间，转染效率最高。血清在本实验室条件下并不影响转染效率。结论：实验获得的优化条件可以明显提高脂质体对肿瘤细胞的转染效率，可作为有关研究或应用的参考。Objective： To confirm the optimized factors affecting the transfection efficiency of liposome to improve transfection efficiency for relevant researches. Method： Using green fluorescent protein as reporter gene, the recombinant plasmid pU6H1 - GFP - FAK was introduced into Caco - 2 cells with Lipofectamine 2000. The following conditions were optimized： cell density, DNA amount, ratio of liposorae and DNA, the forming time of liposome - DNA complex, incubation time of cell with liposome complex, serum, and the times of cell passage. Result： The highest transfection efficiency was achieved with the following optimized conditions： at passage 2 - 5, 2 ×10^5 cells, 4μg DNA, 2.5：1 ratio for liposome and DNA, 30 min for complex formation, and 6h for the incubation. Serum had no effect on the efficiency under these conditions. Conclusion： The liposome complex transfection efficiency can be significantly enhanced under the optimized conditions, and they may be used as the reference for other labs.

关 键 词：脂质体 细胞转染 转染效率 肿瘤细胞 

分 类 号：Q782[生物学—分子生物学] R735.35[医药卫生—肿瘤]