鸦胆子油乳联合siRNA-ERCC1对肺腺癌 A549/DDP细胞的耐药逆转作用

目的:探讨靶向切除修复交叉互补基因1(ERCC1)的小干扰RNA片段(small interfering RNA,siRNA)和鸦胆子油乳联合应用逆转人肺腺癌耐药细胞A549/DDP的效果。方法:设计并合成针对ERCC1基因对应序列的siRNA,采用转染试剂Lipofectamine 2000转染人肺腺癌耐药细胞A549/DDP;将实验组细胞分为:空白对照组、鸦胆子油处理组、siRNA处理组及鸦胆子油联合siRNA处理组,利用RT-PCR检测ERCC1 mRNA和Western blot检测ERCC1蛋白质的表达;MTT法检测A549/DDP细胞对顺铂的耐药逆转效果。结果:siRNA、鸦胆子油、鸦胆子油和siRNA联合应用均能降低ERCC1 mRNA和蛋白质的表达,对顺铂的敏感性明显恢复,耐药倍数分别降至9.72,8.89,7.59,6.83,鸦胆子油和siRNA联合应用效果明显提高(P<0.05)。结论:siRNA、鸦胆子油能有效地逆转人肺腺癌耐药细胞A549/DDP的多药耐药,鸦胆子油和siRNA联合应用效果明显加强。

Reversal Effect of Brucea javanica Oil Emulsion Combined siRNA-ERCC1 on Human Lung Adenocarcinoma A549/DDP Cells

Objective:To study the effect of small interfering RNAs(siRNAs) combined with Brucea javanica oil emulsion targeted ERCC1 Gene on reverting platinum resistance of human lung adenocarcinoma cell line A549/DDP. Method: The siRNAs which targeted the same sequences of ERCC1 gene were designed and synthesized.Human lung adenocarcinoma cell line A549/DDP was cultured and tranfected with siRNAs by Lipofectin2000.Experimental cell grouping:A549/DDP(Control),Brucea-treated group,siRNA-treated group,Brucea and siRNA treated group. ERCC1mRNA was assayed by RT-PCT,and the protein expression was detected by Western blotting.The efficiency of A549/DDP to cisplatin was determined by MTT method. Result: The expression of ERCC1mRNA and protein decreased significantly after transfection of siRNAs,B. javanica oil and B. javanica oil+siRNAs.The efficiency of A549/DDP to cisplatin reversed significantly. The drug-resistant multiple droped back to 9.72,8.89,7.59,6.83.The reversing effect increased significantly by siRNAs combined with B. javanica oil. Conclusion: Both siRNAs and B. javanica oil can reverse platinum resistance of human lung adenocarcinoma cell line A549/DDP.The inhibition effect increases significantly by siRNAs combined with B. javanica oil emulsion.