不同RNAi对肝癌耐药细胞MDR1基因表达抑制作用比较  

COMPARISON BETWEEN THE INHIBIT FUNCTION OF THE DIFFERENT RNAI IN THE EXPRESSION OF MDR1 GENE IN HEPATOCELLULAR CARCINOMA CELLS

作　　者：秦川[1] 杨献军[1] 杨瑞[1] 

机构地区：[1]河南新乡医学院分子生物学研究室,新乡453003

出　　处：《现代预防医学》2009年第24期4652-4654,共3页Modern Preventive Medicine

基　　金：河南省自然科学基金资助项目(0611042300)

摘　　要：[目的]探讨应用RNAi技术抑制肝癌耐药细胞MDR1基因表达的抑制作用以及对P-糖蛋白(P-gp)表达和功能的抑制作用。[方法]根据MDR1基因序列,设计并体外转录合成2条siRNA(smallinterferingRNA),用脂质体转染将其导入细胞内。应用MTT检测转染后癌细胞生长增殖情况,用MTT法测定细胞对化疗药物阿霉素(ADM)的敏感性,流式细胞仪检测细胞膜表面P-糖蛋白(P-gp)表达和细胞内罗丹明(Rhdaming123,Rh123)的潴留。[结果]转染sh-MDR1-1和sh-MDR1-2可显著抑制两株耐药细胞MDR1mRNA和P-gp的表达,细胞内的Rh123稳态积累量均明显增高;第1条序列更能有效的抑制MDR1基因表达。[结论]sh-MDR1-1特异性siRNA可更强的抑制肝癌耐药细胞MDR1基因表达。[Objective] To investigate the inhibitive effects of RNA interference（RNAi）on the expression of MDR1 gene in hepatocellular carcinoma cells.[Methods] Two siRNA which targeted MDR1 gene were designed and synthesized by in vitro transcription,and transfered into human hepatocellular carcinoma SMMC-772 cells using liposome transfection reagents.Drug sensitivity was measured by MTT assay,and P-gp expression and intracellular Rh123 accumulation were determined by flow cytometry（FCM）.[Results] sh-MDR1-1 and sh-MDR1-2 could suppress the P-gp expression in human hepatocellular carcinoma SMMC-772 cells.Drug sensitivity was increased significantly after sh-MDR1-1 transferring in SMMC-7721 cells.The level of P-gp expression was significantly reduced.The intracellular accumulation of Rh123 was increased greatly after treatment in cells.The sh-MDR1-1 was more effective in the suppression of MDR1.[Conclusion] sh-MDR1-1 can suppress the expres-sions of MDR1 and P-gp in human hepatocellular carcinoma SMMC-772 cells.

关 键 词：RNA干扰 MDR1基因 肝癌细胞 多药耐药 

分 类 号：R735.7[医药卫生—肿瘤]