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

目的：探讨绿色荧光蛋白(green fluorescence protein，GFP)特异的小干扰RNA(small interference RNA，siRNA)对肺肿瘤细胞A549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siRNA GFP-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。

Specific suppression of GFP gene expression by siRNA of GFP in lung carcinoma cell A549

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.