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

Inhibition of RSK4 gene expression in the breast cancer cell line MCF-7 using lentiviral vector-mediated shRNA interference

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,72h开始检测到GFP荧光表达,90h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%和80.1%。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 RSKd 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.