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中国癌症防治杂志

中国癌症防治杂志 ›› 2013, Vol. 5 ›› Issue (1): 20-23.doi: 10.3969/j.issn.1674-5671.2013.01.05

• 基础研究 • 上一篇    下一篇

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

  

  1. 广西医科大学附属肿瘤医院乳腺外科
  • 出版日期:2013-03-25 发布日期:2013-04-07
  • 通讯作者: 刘剑仑 E-mail:jianlunliu@hotmail.com
  • 基金资助:

    国家自然科学基金资助项目（30960427）；广西自然科学基金资助项目（桂科自0991231）

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

  • Online:2013-03-25 Published:2013-04-07

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被引次数

8

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％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。

关键词: 乳腺肿瘤, 核糖体S6蛋白激酶4, shRNA干扰, 慢病毒载体

Abstract:  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. Conclusion 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.

Key words:  Breast neoplasms, RSK4, shRNA interference, Lentiviral