| 引用本文: | 沈雁，李瑞珺，范达，李谦，涂家生.特异性抑制环氧合酶-2表达的siRNA筛选[J].中国现代应用药学,2012,29(5):377-380. |
| SHEN Yan, LI Ruijun, FAN Da, LI Qian, TU Jiasheng.Selection of Specific siRNA Inhibiting the Expression of COX-2[J].Chin J Mod Appl Pharm(中国现代应用药学),2012,29(5):377-380. |
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| 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。 |
| 关键词: 环氧合酶-2 siRNA RT-PCR 转染 |
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| 基金项目: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) |
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| Selection of Specific siRNA Inhibiting the Expression of COX-2 |
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SHEN Yan, LI Ruijun, FAN Da, LI Qian, TU Jiasheng1,2
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1.China Pharmaceutical University, a.Department of Pharmaceutics;2.b.School of Life Science and Technology, Nanjing 210009, China
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| 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. |
| Key words: COX-2 siRNA RT-PCR transfection |
