论文部分内容阅读
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 evaluate and compare the transfection efficiency and toxicity of A549 cells with two transfection reagents Lipofectamine 2000 and jet PRIMETM. Methods The plasmid pEGFP-N1 was transfected into A549 cells using Lipofectamine 2000 and jet PRIMETM respectively. The expression of enhanced green fluorescent protein (EGFP) was observed under a fluorescence microscope 24h after transfection. The transfection efficiency, The toxicity of the two transfection reagents to A549 cells was tested by [3- (4.5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide]. Results When the ratio of plasmid to Lipofectamine 2000 was changed from 1: 2 to 1: 3.5, the transfection efficiency of Lipofectamine 2000 did not change obviously. The cytotoxicity increased with the increase of the amount of Li-pofectamine 2000. The highest transfection rate was 50.6 ± 4.9% and the cell survival rate was (89.2 ± 9.1)% when the ratio of plasmid to jet PRIMETM was 2000 (μg / μl) The transfection efficiency of jetPRIMETM changed obviously, while the change of cytotoxicity was not obvious. The transfection rate was (30.6 ± 2.8)% when the ratio of plasmid to jet PRIMETM (μg / μl) was 1: 2 The survival rate was (100.6 ± 4.8)%. The maximum transfection rate and the corresponding cell survival rate were statistically different (P <0.01, P <0.05). Conclusions Lipofectamine 2000 transfection of A549 cells is more efficient than jetPRIMETM and cytotoxicity is stronger than jetPRIMETM.