两种转染试剂转染重组质粒至Caco-2细胞转染效率的比较

Comparison of efficiency of two transfection reagents in transfecting recombinant plasmid into Caco-2 cells

目的比较两种转染试剂Lipofectamine LTX&PLUS和罗氏X-tremeGENE HP DNA Transfection Reagent转染重组质粒至Caco-2细胞的转染效率。方法分别采用Lipofectamine LTX&PLUS转染试剂与罗氏X-treme GENE HP转染试剂转染Caco-2细胞,以荧光素酶(Luc)和绿色荧光蛋白(GFP)作为报告基因,利用荧光信号检测仪检测荧光信号,Lipofectamine LTX和质粒的比值分别为4∶1,3.5∶1,2∶1,罗氏X-HP与质粒的比值分别为3∶1,2∶1,1∶1,对转染试剂和质粒DNA之间的比值进行优化,然后比较两种转染试剂的转染效率;荧光倒置显微镜观察转染效果。结果对于Caco-2细胞,两种转染试剂均未影响细胞的正常形态,细胞毒性小。罗氏X-HP转染试剂与DNA质粒的比例是3∶1时转染效率较高(P<0.05),LTX转染试剂与DNA质粒的比例为3.5∶1时转染条件最佳(P<0.05)。比较两者的转染效率,罗氏转染试剂(Firefly RLU/Renilla RLU=0.667)高于LTX转染试剂(Firefly RLU/Renilla RLU=0.502)(P<0.05)。结论不同的转染试剂对相同细胞的转染效率不同。相比较而言,罗氏转染试剂对于Caco-2细胞的转染更为合适。

Objective To compare the transfection efficiency of the two transfection reagents used in transfection of recombinant plasmid into Caco-2 cells.Methods Caco-2 cells were transfected with recombinant plasmids labeled with luciferase（LUC） and green fluorescent protein（GFP） as the reporter genes using Lipofectamine LTX ＆ PLUS transfection reagent and Roche X-tremeGENE HP DNA transfection reagent as the transfection reagents. The ratio of Lipofectamine LTX and plasmid was 4 ∶ 1, 3.5 ∶ 1, 2 ∶ 1 respectively, and the ratio of Roche X-HP to plasmid was 3 ∶ 1,2 ∶ 1 and 1 ∶ 1. The dosage of the transfection reagent and plasmid DNA was optimized and the transfection efficiency of the two transfection reagents was compared using the fluorescent signal detector and fluorescent inverted microscope.Results Neither of the two transfection reagents affected the morphology of the cells, indicating their cytotoxicity was slight. The best proportion of LTX transfection reagent and DNA plasmid was 3.5 ∶ 1（ P 〈0.05）, and the best ratio of Roche X-HP transfection reagent and DNA plasmid was 3 ∶ 1（ P 〈0.05）. In addition, the transfection efficiency of Roche transfection reagent（Firefly RLU/Renilla RLU=0.667） was higher than that of LTX transfection reagent（Firefly RLU/Renilla RLU=0.502）（ P 〈0.05）.Conclusion The transfection efficiency of different transfection reagents varies in the same cells. Roche transfection reagent is more suitable for the transfection of Caco-2 cells.