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

    • 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50 μmol·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。

       

      Abstract: 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(P<0.01). CONCLUSION siRNA954 can inhibit COX-2 expression effectively. It plays an important role in the treatment of tumor.

       

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