筛选介导化学合成siRNA转染原代肝癌细胞的最佳试剂

Transfection agents screening for chemosynthesis siRNA transfection to primary liver cancer cells

背景:理想的细胞转染试剂应具有高效安全的特点。目的:筛选能够高效介导化学合成siRNA转染原代肝癌细胞的最佳转染试剂。方法:应用Lipofectamine RNAiMAX,Lipofectamine2000和DharmaFECT1转染试剂介导FAM-siRNA和多药耐药基因MDR1siRNA转染原代肝癌细胞,分别于转染6和48h后应用流式细胞仪和实时荧光定量PCR检测转染效率,然后用MTT法检测3种转染试剂处理原代肝癌细胞24h后的细胞毒性。结果与结论:对于Lipofectamine RNAiMAX,Lipofectamine2000和DharmaFECT1转染试剂介导的FAM-siRNA和MDR1siRNA转染,流式细胞仪和实时荧光定量PCR仪检测出RNAiMAX转染效率最高(P<0.05),分别为70.3%和71.5%。MTT法检测结果表明RNAiMAX对原代肝癌细胞没有表现出细胞毒性。结果提示,由于RNAiMAX介导的FAM-siRNA和MDR1siRNA转染的效率最高,并且对细胞的毒性最小,所以RNAiMAX是最适合介导化学合成siRNA转染原代肝癌细胞的转染试剂。

BACKGROUND：The ideal transfection agent should efficient and low-toxic.OBJECTIVE：To screen transfection agent which efficiently transfect chemosynthesis siRNA to primary liver cancer cells.METHODS：FAM-siRNA and MDR1 siRNA was transfected to primary liver cancer cells by Lipofectamine RNAiMAX,Lipofectamine 2000 and DharmaFECT1.The transfection efficiency was evaluated by flow cytometer and real time-PCR at 6 and 48 hours after transfection.Moreover,the cytotoxicity of transfection agents in primary liver cancer cells was tested by MTT method.RESULTS AND CONCLUSIONS：The results of flow cytometer and real time-PCR indicated that,the transfection efficiency of siRNA tranfection to primary liver cancer cells mediated by RNAiMAX was highest （P0.05）,which was 70.3% and 71.5%,respectively.The cytotoxicity of RNAiMAX in primary liver cancer cells did not show in MTT detection （P0.05）.RNAiMAX was suitable to transfect siRNA to primary liver cancer cells because the efficiency of siRNA tranfection to primary liver cancer cells mediated by RNAiMAX was the highest and the cytotoxicity of RNAiMAX in primary liver cancer cells was the lowest.