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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 screen small interfering RNA (siRNA) sequences that can effectively inhibit cyclooxygenase-2 (COX-2) expression and to observe its inhibitory effect on COX-2 expression. Methods Three pairs (siRNA24, siRNA954, siRNA1553) of COX-2 and one FAM fluorescently-labeled negative control siRNA (called HK) were designed. Human gastric cancer cell line SGC-7901 was transfected with Lipofectamine2000. Meanwhile, the content of COX-2 in cells was detected by RT-PCR and Western blot, and β-actin protein was used as internal reference. Results The maximum transfection efficiency was achieved when the concentration of Lipofectamine 2000 was 50 μmol·L -1. Compared with other groups, the expression of COX-2 mRNA was significantly decreased in human gastric cancer cells transfected with siRNA954 (P <0.01). Conclusion The siRNA against COX-2 was successfully designed and siRNA954 was screened out. It can effectively inhibit the expression of COX-2 in human gastric cancer cells and lay a foundation for the treatment of cancer and its clinical application.