Lipofectamine 2000和jetPRIME^(TM)对A549细胞转染效率及毒性的比较研究

Comparative Study of Transfection Efficiency and Cytotoxicity of DNA into A549 Cells by Two Transfection Reagents:Lipofectamine 2000 and JetPRIMETM

目的评价及比较两种转染试剂Lipofectamine 2000和jetPRIMETM对A549细胞的转染效率及毒性。方法分别利用Lipofectamine 2000和jetPRIMETM将pEGFP-N1质粒转染A549细胞,转染24h后荧光显微镜观察增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)的表达,计算转染效率,四甲基偶氮唑盐比色法[3-(4.5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide,MTT]检测两种转染试剂对A549细胞的毒性。结果质粒与Lipofectamine 2000比例(μg/μl)在1∶2至1∶3.5之间变化时,Lipofectamine 2000转染效率变化不明显,细胞毒性随着Li-pofectamine 2000用量增多而增大,质粒与Lipofectamine 2000比例(μg/μl)为1∶2.5时转染率最高,达(50.6±4.9)%,细胞存活率为(89.2±9.1)%;质粒与jetPRIMETM比例(μg/μl)在1∶1至1∶4之间变化时,jetPRIMETM转染效率变化明显,而细胞毒性变化不明显,质粒与jetPRIMETM比例(μg/μl)为1∶2时转染率最高,为(30.6±2.8)%,细胞存活率为(100.6±4.8)%;两者最大转染率及相应的细胞存活率均有统计学差异(P<0.01,P<0.05)。结论 Lipofectamine 2000转染A549细胞的效率高于jetPRIMETM,细胞毒性亦强于jetPRIMETM。

Objective To compare and evaluate the transfection efficiency and cytotoxicity of DNA into A549 cells by two transfection reagents： Lipofectamine 2000 and jetPRIMETM.Methods Lipofectamine 2000 and jetPRIMETM were employed for the transfection of plasmid pEGFP-N1,a plasmid that express enhanced green fluorescent protein（EGFP）,into A549 cells.The expression of EGFP was observed 24 hours after transfection by fluorescent microscope and the transfection efficiency was calculated.3-（4.5-dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium bromide（MTT） assay was used to observe the cytotoxicity of the transfection reagents.Results The transfection efficiency of Lipofectamine 2000 didn＇t change significantly when the ratio of plasmid/Lipofectamine 2000（μg/μl） was between 1∶2 and 1∶3.5,but the cytotoxicity of Lipofectamine 2000 was increased along with the increasing dosage of Lipofectamine 2000.The highest transfection efficiency was（50.6±4.9）% when the ratio of plasmid/Lipofectamine 2000（μg/μl） was 1∶2.5 and at the same time,the cell survival rate was（89.2±9.1）%.The transfection efficiency of jetPRIMETM changed significantly when the ratio of plasmid/jetPRIMETM（μg/μl） was between 1∶1 and 1∶4,but the cytotoxicity of jetPRIMETM didn＇t change along with the increasing dosage of jetPRIMETM.The highest transfection efficiency was（30.6±2.8）% when the ratio of plasmid/ jetPRIMETM（μg/μl） was 1∶2,and at the same time,the cell survival rate was（100.6±4.8）%.The transfection efficiency and cytotoxicity of two transfection reagents were significantly different（P0.01,P0.05）.Conclusion Lipofectamine 2000 has higher transfection efficiency and cytotoxicity than jetPRIMETM.