小干扰RNA特异性抑制肺肿瘤细胞A549中绿色荧光蛋白基因表达
Specific suppression of GFP gene expression by siRNA of GFP in lung carcinoma cell A549
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摘要: 目的: 探讨绿色荧光蛋白(green fluorescence protein,GFP)特异的小干扰RNA (smallinterference RNA,siRNA)对肺肿瘤细胞A54 9中的GFP基因表达的抑制作用,建立以真核表达载体释放生成特异基因siRNA的方法。方法: 构建mU6启动子驱动的含有GFP特异siRNA的真核表达质粒,转染A549肺肿瘤细胞,观察共转染时不同剂量siRNA对GFP的抑制效应。结果: 针对GFP核酸序列设计的特异性siRNAGFP-siRNA可有效抑制其表达,并当比例为1∶1时效果最佳,而空质粒载体及无关RNAi无此效应。结论: 以mU6为启动子的含特异RNAi的质粒可作为抑制特异基因表达的一种新的技术手段,并且为研究基因功能的一条有效途径。Abstract: Objective: It was designed to investigate whether small interfering RNA （siRNA） of green fluorescence protein （GFP） can inhibit the expression of GFP in lung carcinoma cell A549. Methods: A plasmid included mU6 promoter and siRNA of GFP was designed and constructed. The plasmid was transfected into lung carcinoma cell A549. GFP-siRNA effect of interference to GFP is observed and evaluate in A549 cell. Results: GFP-RNAi can inhibit specific expression of GFP gene in lung carcinoma A549 cell line and mU6 plasmid, no-related-siRNA have no such specific effect. Conclusion: These observations may open a path forward the use of siRNA as a new tool for researching gene function in cancer cell line.
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