FuGNE6与pGPu6/GFP/neo-GPC3-iRNA质粒不同比率对HepG2细胞转染效率的影响  

Effect of different ratios of FuGNE6 to pGPu6/GFP/neo-GPC3-iRNA on transfection efficiency in Hep G2 cells

作　　者：姜菲菲[1] 赵焕英[2] 隋雷鸣[2] 绳波[1] 徐萌[1] 李莉[1] 

机构地区：[1]首都医科大学北京佑安医院临床检验中心,北京100069 [2]首都医科大学医学实验与测试中心

出　　处：《山西医科大学学报》2015年第1期25-27,100,共4页Journal of Shanxi Medical University

基　　金：首都医科大学校基金技术类资助项目(2014JS08;2014JS06);首都医科大学基础临床科研合作基金资助项目(13JL28)

摘　　要：目的探讨转染试剂FuGNE6与pGPu6/GFP/neo-GPC3-iRNA质粒不同比率对HepG2细胞转染效率的影响,并寻找最佳的转染比例。方法采用转染试剂FuGNE6(以6μl为例)与GPC3-iRNA质粒(μg)比率分别为6∶0.5,6∶1,6∶1.5,6∶2,6∶4,6∶6及GPC3-iRNA量固定1μg(以此为例),转染试剂与GPC3-iRNA质粒(μg)比率分别为2∶1,4∶1,6∶1,8∶1,10∶1进行转染。分别用荧光显微镜、流式细胞仪及PI染色实验观察细胞转染效率及48h细胞存活率。结果FuGNE6固定为6μl时,质粒用量从0.5-2μg转化效率逐步增高,超过2μg后转染效率反而大大降低,各比例组在48h细胞存活率都在95%以上;质粒量固定为1μg时,随转染试剂增加转化效率不断增大,超过6∶1时,随转染试剂增加,48h细胞存活率也逐渐下降。结论FuGNE6与GPC3-iRNA比例为6∶1.5时转化效率最大,且细胞存活率高。Objective To explore the effects of different ratios of FuGNE6 to pGPu6/GFP/neo-GPC3-iRNA （transfection vectors）on transfection efficiency in HepG2 cells, and to find out the optimized ratio. Methods GPC3-iRNA was transfected with FuGNE6 and the transfection efficiency was compared at different ratios of GPC3-iRNA to transfection reagent （FuGNE6）. The different amounts of GPC3-iRNA was added to 6 trl of FuGNE6 at different FuGNE6-to-GPC3-iRNA ratios of 6： 0.5, 6： 1, 6： 1.5, 6： 2, 6： 4, 6： 6,respectively. Additionally, FuGNE6 was added to 1 μg of DNA at different FuGNE6-to-GPC3-iRNA ratios of 2： 1,4： 1,6： 1,8： 1, 10： 1, respectively. Fluorescence microscope, flow cytometer and PI staining were used to detect the transfection efficiency and the cell survival rate at 48 h after transfection. Results When the amount of FuGNE6 was 6 μl, the transfection efficiency gradually increased in the range of 0. 5 - 2 μg GPC3-iRNA, and the transfection efficiency was significantly reduced when the amount of GPC3-iRNA was higher than 2 μg. The cell survival rate was more than 95% in all groups at 48 h after transfection. At the same GPC3-iRNA concentration （ 1 μg）, the transfection efficiency gradually rose with the increase of transfection reagent. When the FuGNE6-to-DNA ratio was over 6： 1, the cell survival rate was gradually decreased at 48 h after transfection. Conclusion The transfection efficiency and the cell survival rate are the highest when the FuGNE6-to-DNA ratio is 6： 1.5.

关 键 词：转染 FuGENE6 HEPG2细胞 

分 类 号：Q81[生物学—生物工程]