基础医学与临床 ›› 2012, Vol. 32 ›› Issue (11): 1288-1292.

• 研究论文 • 上一篇    下一篇

SiRNA沉默Jab1表达抑制肝癌细胞增殖

程晓刚1,叶琬2,张绍兰1,陈玮1,金虹1,徐艳1,李晋川1   

  1. 1. 成都医学院
    2. 成都蓉生药业有限公司
  • 收稿日期:2012-01-17 修回日期:2012-03-28 出版日期:2012-11-05 发布日期:2012-10-19
  • 通讯作者: 程晓刚 E-mail:cxg92@sina.com
  • 基金资助:
    四川省教育厅应用基础研究项目

SiRNA Silencing JAB1 Expression Inhibit Hepatoma Cells Proliferation

  • Received:2012-01-17 Revised:2012-03-28 Online:2012-11-05 Published:2012-10-19
  • Contact: 程晓刚 Cheng Xiao-Gang E-mail:cxg92@sina.com

摘要: 目的 观察沉默Jab1基因表达对人肝癌细胞HepG-2增殖的影响。方法 利用RNA干扰技术构建PAVU6-JAB1 siRNA干扰质粒,将其转染HepG-2细胞,采用Real-time PCR、Western blot等方法对Jab1表达进行检测,WST-8、流式细胞分析对细胞增殖的影响。结果 成功构建PAVU6- JAB1 siRNA干扰质粒,转染HepG-2细胞后Jab1蛋白表达量较对照组明显降低,P27kip1蛋白表达量明显升高 (P<0.01)。转染96 h后能显著抑制细胞增殖活性,由空载体转染组的99.6%?1.4%降至70.8%?1.6%。结论 构建靶向Jab1基因的干扰质粒能下调Jab1基因的表达,上调P27kip1蛋白水平,并抑制HepG-2细胞在体外的增殖活性。

关键词: JAB1, RNA干扰, 肝癌细胞, 增殖

Abstract: Objective Observe the effect of silencing JAB1 gene expression on human hepatoma cells proliferation. Methods By means of the RNA interference (RNAi) technique, the pAVU6-JAB1 siRNA interference vectors were constructed and transfected into hepatoma cell line HepG-2. The Jab1 expression was measured using real-time PCR and weatern-blot methods. The cells proliferation was measured using WST-8 methods. Results 96 hours after transfection of the PAVU6-JAB1 siRNA vectors, the proliferation of hepatoma cell line were inhibited from 99.6?1.4%(empty vector transfection group) to 70.8?1.6%(experimental group). The Jab1 expression was also inhibited,whereas the P27kip1 expression level was significantly elevated. Conclusion Inhibition of the expression of the JAB1 protein in hepatoma cells could inhibit hepatoma cell proliferation to some extent.

Key words: JAB1, RNA interference, hepatoma cell, proliferation

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