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目的:探索siRNA对恶性黑素瘤LiBr细胞livin基因表达的影响及剂量和时间效应。方法:将化学合成的3条靶向livin基因的siRNA序列转染至人恶性黑素瘤LiBr细胞,分别在mRNA和蛋白水平应用real-time RT-PCR和Westernblot检测其表达变化以确定沉默效应最好的序列并观察其下调livin表达的剂量和时间效应。结果:其中1条siRNA序列显著下调livin mRNA和蛋白表达,并且呈剂量和时间依赖关系;100nmol/L可达到最大沉默效应,其下调livin mRNA和蛋白最大效应分别出现在转染后48h和72h。结论:应用siRNA技术靶向livin mRNA790-808位点可有效地敲低LiBr细胞livin的表达。
Objective: To investigate the effect of siRNA on livin gene expression in LiBr cells and its dose and time effect. Methods: Three chemically synthesized livin siRNA sequences were transfected into human malignant melanoma LiBr cells. The expression of livin mRNA was detected by real-time RT-PCR and Western blot respectively to determine the effect of silencing Good sequence and observe its dose and time effect of down-regulated livin expression. Results: One siRNA sequence significantly down-regulated the expression of livin mRNA and protein in a time-and dose-dependent manner. 100 nmol / L reached the maximum silencing effect, and the maximum effect of livin mRNA and protein down-regulation occurred at 48 and 72 h after transfection. Conclusion: siRNA targeting livin mRNA790-808 can effectively knock down livin expression in LiBr cells.