【摘要】 目的观察RNA干扰沉默膜联蛋白-1(Annexin-1)基因表达技术对膀胱癌T24细胞Annexin-1基因表达和顺铂耐药的影响。方法针对Annexin-1基因不同部位设计并化学合成3对不同的靶向小分子干扰RNA(siRNA),脂质体介导瞬时转染膀胱癌T24细胞,转染后应用半定量RT-PCR和Western印迹检测Annexin-1mRNA和蛋白的改变,用MTT法检测沉默Annexin-1表达在膀胱癌T24细胞对顺铂敏感性的变化。结果转染siRNA后膀胱癌T24细胞Annexin-1的mRNA和蛋白水平显著下降(P<0.05),增强细胞对顺铂的敏感性(P<0.05)。结论 RNA干扰Annexin-1基因后其mRNA和蛋白水平显著下降,并且增强T24细胞对顺铂的敏感性。更多还原

【Abstract】 Objective To explore the effects of annexin-1 gene silencing by small interfering RNA(siRNA)on bladder cancer cells T24 and the sensitivity change of DDP.Methods Three siRNA targeting three different domains of annexin-1 gene were designed,synthesized,and transfected into bladder cancer T24 cells by lipofectamineTM2000.After transfection,expression of annexin-1 was detected by RT-PCR and West-blotting.The cellular sensitivity to DDP was evaluated by MTT assay.Results Compared with the negative control(siRNA-NC)groups,transfection of three targeting siRNA in T24 cells decreased the expression of annexin-1 mRNA and protein.Annexin-1 gene silencing increased the sensitivity of T24 cells to DDP.Conclusions The chemically synthesized specific siRNA targeting annexin-1 could effectively reduce the expression of annexin-1 gene,increase the sensitivity of T24 cells to DDP.更多还原