特异性抑制环氧合酶-2表达的siRNA筛选  

Selection of Specific siRNA Inhibiting the Expression of COX-2

作　　者：沈雁[1] 李瑞珺[1] 范达[1] 李谦[2] 涂家生[1] 

机构地区：[1]中国药科大学药剂学教研室,南京210009 [2]中国药科大学生命科学院,南京210009

出　　处：《中国现代应用药学》2012年第5期377-380,共4页Chinese Journal of Modern Applied Pharmacy

基　　金：中央高校基本科研业务费专项基金资助项目(JKQ2011018);国家自然科学基金资助项目(81072588)

摘　　要：目的筛选有效抑制环氧合酶-2(COX-2)表达的小干扰RNA(siRNA)序列,并观察其对细胞中COX-2表达的抑制作用。方法设计针对COX-2的3对(siRNA24,siRNA954,siRNA1553)及一条FAM荧光标记的阴性对照siRNA(称为HK)。在Lipofectamine2000介导下转染人胃癌细胞系SGC-7901。同时采用RT-PCR法和Western blot方法检测细胞中COX-2含量,以β-actin蛋白做内参照。结果当Lipofectamine2000浓度在50μmol.L 1时能实现最大转染效果。与其他各组相比,转染了siRNA954的人胃癌细胞中,编码COX-2 mRNA的含量明显减少(P<0.01)。结论成功设计了针对COX-2的siRNA,并从中筛选出siRNA954,能够有效抑制人胃癌细胞COX-2表达,为肿瘤治疗及其临床应用奠定了基础。OBJECTIVE To select the siRNA which could most effectively inhibit the expression of COX-2,and to observe the change of COX-2 expression in cells.METHODS Three siRNAs were designed（siRNA24,siRNA954,siRNA1553） and one FAM fluorescence-labeled siRNA was used as a negative control（HK）.Detect the effect of inhibition by RT-PCR and Western blot after transfecting human gastric cancer cell line SGC-7901.RESULTS The 50 μmol·L 1 Lipofectamine2000 had highest transfection effiency.The expression of human COX-2 gene was suppressed specific and effectively by siRNA954,compared with other groups（P0.01）.CONCLUSION siRNA954 can inhibit COX-2 expression effectively.It plays an important role in the treatment of tumor.

关 键 词：环氧合酶-2 SIRNA RT-PCR 转染 

分 类 号：R963[医药卫生—微生物与生化药学]