转铁蛋白增强脂质体介导基因对肝癌细胞的转染和表达  

Enhancement of Liposome-mediated Gene Transfection and Expression in Hepatoma Cells by Transferrin

作　　者：王克敏[1] 赵宇[1] 夏爱娣[1] 陈诗书[1] 

机构地区：[1]上海第二医科大学生化教研室分子生物学实验室,200025

出　　处：《胃肠病学》2001年第3期135-138,共4页Chinese Journal of Gastroenterology

基　　金：闰家自然科学基金重点课题(No39730440);上海市免疫研究所资助

摘　　要：目的：检测转铁蛋白能否增强脂持体介导外源基因对肝癌细胞的转染和表达。方法：应用本滓验室构建的含小 鼠一细胞介素（ｍｌＬ）－１２的真核表达质粒ｐｃＤＮＡ３ ｍＩＬ－１２和绿色荧光蛋白（ＧＦＰ）真核表达质粒ｐｃＤＮＡ３－ ＥＧＦＰ， 观察转铁蛋白增强阳离子脂质体ＬｉｐｏｆｅｃｔＡＭＩＮＥ介导上述两种质粒ＤＮＡ对小鼠和不同人肝癌细胞株的转染及 表达效率。结果：ＬｉｐｏｆｅｃｔＡＭＩＮＥ加转铁蛋白介导ｐｃＤＮＡ３／ＥＧＲＰ对小鼠肝癌细胞株Ｈｅｐｌ－６的转染效率由２％－ ５％提高至２０％，对４种不同人肝癌细胞株的转染效率由１５％。提高至６５％～７５％；但对小鼠肝癌细胞株ＭＭ４５ＴＬｉ 的转染效率并不增加。转铁蛋白可使ＬｉｐｏｆｅｃｔＡＭＩＮＥ介导ｐｃＤＮＡ３ｍＩＬ－１２在Ｈｅｐｌ－６中的表达量增强１８倍。结论： 转铁蛋白能够与ＬｉｐｏｆｅｃｔＡＭＩＮＥ和质粒ＤＮＡ形成复合物，显著增强脂质体介导外源基因对肝癌细胞的转染和表 达，但对不同种肝癌细肥株的增强效率不同。Background Aims: To detect the enhancement of liposome-mediated gene transfection and ex- pression in hepatoma cells by transferrin. Methods: Eukaryotic expressing vector containing murine interleukin (mIL)-12 and green fluorescent protein (GFP), i.e. pcDNA3 mIL-12 and pcDNA3 EGFP con- structed in our laboratory were used to detect the enhancement of lipofectAMINE-mediated gene transfection and expression in different hepatoma cell lines of murine and human by transferrin. Re- sults: The transfection rate of murine hepatoma cell line Hepl-6 increased from 2%~5% to 20% by transferrin-lipofectAMlNE-pcDNA3 EGFP complex, and from 15% to 65%~75% in four human hepato- ma cell lines, but did not increase in murine hepatoma cell line MM45T.Li. Transferrin increased mIL- 12 expression in Hepl-6 by lipofcctAMlNE-mediated pcDNA3 mIL-12 about 18 folds over lipofect- AMINE transfection alone. Conclusions: The transferrin-lipofectAMlNE-plasmid DNA complex can be formed. Transferrin can increase the transfection and expression rates of foreign gene in hepatoma cells significantly as mediated by lipofectAMlNE. but the enhancement rates of different hepatoma cell lines are different.

关 键 词：转铁蛋白 白细胞介素12 脂质体 肝肿瘤 细胞系 绿色荧光蛋白 

分 类 号：R735.7[医药卫生—肿瘤]