靶向Bel-7402/ADM细胞MDR1基因高效siRNA的构建与鉴定  

Construction and Identification of SiRNA Targeting to MDRl of Bel-7402/ADM

作　　者：盛龙[1] 熊茂明[1] 李聪[1] 孟翔凌[1] 

机构地区：[1]安徽医科大学第一附属医院普外科,合肥230022

出　　处：《肝胆外科杂志》2012年第4期304-307,共4页Journal of Hepatobiliary Surgery

基　　金：安徽高校省级自然科学研究(重点)项目(KJ 2011A155);安徽省卫生厅医学科学研究基金(2010B007)

摘　　要：目的构建并鉴定靶向人肝癌多药耐药细胞亚系Bel-7402/ADM细胞MDRl基因的高效siRNA,为逆转肝癌多药耐药提供新的分子靶点。方法设计并化学合成4条靶向MDRl的siRNAs(mdrlsi-1、mdrlsi-2、mdrlsi-3和mdrlsi-4),通过Li-pofectamineTM2000介导转染Bel-7402/ADM细胞株,用RT-PCR检测Bel-7402/ADM细胞MDRl mRNA表达、western blot检测P-gp的表达,综合评价这4条siRNAs的沉默效果。结果经siRNA干扰后,4条siRNAs均能不同程度逆转Bel-7402/ADM细胞MDR1介导的多药耐药,mdrlsi-1组mRNA和蛋白表达与其他组比较有显著性差异(P<0.05)。结论相比较其他3条siRNAs,mdr1si-1对人肝癌Bel-7402/ADM细胞MDRl介导的耐药逆转效果最好,其靶序列有望成为逆转肝癌多药耐药新的靶点。Objective To explore new molecular targets for the reversal of hepatocellular carcinoma muhidrug resistance （MDR）, efficient siRNA targeting MDR1 gene of Bel-7402/ADM was constructed and identified. Methods Small interfering RNAs （siRNAs） of four（ mdrlsi-1, mdrlsi-2, mdrlsi-3 and mdrlsi-4） targeting MDR1 were firstly designed and processed with chemical syn- thesis. Mediated by LipofectamineTM 2000, the siRNAs was transfected into Bel-7402/ADM cell strains. The mRNA expression of MDR-1 gene was evaluated by RT-PCR. And the product of P-glyeoprotein （P-gp） was examined by Western blot. Based on these as- says, the gene silencing effect of these four siRNAs was comprehensively evaluated. Results With the siRNA interference, four siRNAs reversed MDR in hepatocellular carcinoma mediated by MDR1 to varying degrees. Compared with other groups, mdrl mRNA and P-gp expression in the mdrlsi-1 group presented significant difference （P 〈 0. 05 ）. Conclusion： mdrl si-1 is a promising candidater for the reversal of MDR of hepatocellular carcinoma.

关 键 词：SIRNA RNA干扰 MDR1基因 Bel-7402/ADM细胞株 

分 类 号：R735.7[医药卫生—肿瘤]