慢病毒载体介导shRNA干扰对乳腺癌MCF-7细胞RSK4基因表达的影响

摘要

目的观察慢病毒干扰载体（RSK4-RNAi-LV）转染对低侵袭、低转移潜能的乳腺癌 MCF-7细胞中核糖体 S6蛋白激酶4（ribosomal protein S6 kinase 4，RSK4）基因表达的影响。方法设计合成特异性的 RSK4 shRNA 干扰体 RSK4-RNAi-LV，稳定转染至乳腺癌 MCF-7细胞中，应用实时荧光定量 PCR 法和 Western blot 法检测 RSK4 mRNA 及其蛋白的表达情况。结果稳定转染慢病毒干扰载体至乳腺癌细胞后，72 h 开始检测到 GFP 荧光表达，90 h 后 GFP 荧光表达明显增强，RSK4 mRNA 及其蛋白的表达均被明显抑制，抑制率分别为98.2豫和80.1豫。结论稳定转染 RSK4-RNAi-LV 可有效抑制乳腺癌 MCF-7细胞中 RSK4 mRNA 及其蛋白的表达。%Objective To design a lentiviral vector-based system for exploring the effects of silencing the ribosomal protein S6 kinase 4(RSK4)gene in the human breast cancer cell line MCF-7. Methods A lentiviral RNA interference vector targeting the RSK4 gene was designed and stably transfected into MCF-7 cells.Expression of RSK4 mRNA and protein was analyzed by real-time PCR and Western blotting. Results Stable transfection led to 98.2% inhibition of RSK4 mRNA and 80.1% inhibition of RSK4 protein. Con-clusion Stable transfection of the breast cancer cell line MCF-7 using a lentiviral RNA interference vector can significantly inhibit expression of RSK4 mRNA and protein.