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

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。

Comparative Study of Transfection Efficiency and Cytotoxicity of DNA into A549 Cells by Two Transfection Reagents: Lipofectamine 2000 and 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(P<0.01,P<0.05).Conclusion Lipofectamine 2000 has higher transfection efficiency and cytotoxicity than jetPRIMETM.