中文摘要：

目的比较3种不同转染试剂介导siRNA转染蚊细胞的优缺点,选择最优方法用于转染后细胞存活率检测试验。方法分别用FuGENE HD,X-tremeGENE和siPORT Amine 3种转染试剂介导特定siRNA转染蚊细胞,转染48 h后采用实时荧光定量PCR检测目的基因的表达。比较3种方法的干涉效率,转染试剂对细胞的毒性和不同方法的操作步骤对实验结果的影响。结果 FuGENE HD、X-tremeGENE和siPORT Amine转染后的干涉效率分别为70%、85%和29%;FuGENE HD和siPORT Amine对蚊C6/36抗性细胞毒性较小,X-tremeGENE对蚊细胞毒性较大;FuGENE HD的转染过程需要更换一次细胞培养液,X-tremeGENE的转染过程需要更换2次细胞培养液,siPORT A-mine的转染过程不需要更换培养液。结论转染试剂的转染效果对于实验结果有显著影响。在蚊细胞转染siRNA的实验中,FuGENE HD转染效率尚可,对细胞毒性小,转染过程只需1次换液操作;X-tremeGENE转染效率较高,对蚊细胞毒性较大,有2次换液操作;siPORT Amine毒性小且不需换液操作,但转染效率较低。综合评价：FuGENE HD更适用于转染siRNA后细胞存活率实验。

英文摘要：

Objective To compare the advantages and disadvantages of three different transfection reagents for mediated transfection of siRNA to mosquito cells and choose the most suitable for cell viability assay after transfection.Methods The specific siRNA was transfected into mosquito cells using FuGENEHD,X-tremeGENE,and siPORT Amine transfection reagents.Quantitative Real-Time PCR was performed to access gene expression 48 hr after transfection.The transfection efficiency,cytotoxicity,and use of different reagents were compared.Results Transfection efficiency was 70% for FuGENEHD,85% for X-tremeGENE,and 29% for siPORT Amine.FuGENEHD and siPORT Amine had lower cytotoxicity than did X-tremeGENE.During transfection,FuGENEHD in the culture medium had to be replaced once,X-tremeGENE had to be replaced twice,and siPORT did not have to be replaced.Conclusion The efficiency of transfection reagents significantly affected results.FuGENEHD has moderate transfection efficiency,low cytotoxicity,and was only replaced once during transfection.X-tremeGENE has high transfection efficiency but high cytotoxicity and was replaced twice during transfection.siPORTTM Amine has low cytotoxicity and was not replaced but has low transfection efficiency.