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Effects of small RNA interference to target cyclinE gene on apoptosis and proliferation of HepG2 hepatocellular carcinoma cells
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- Author:
- No author available
- Journal Title:
- WORLD CHINESE JOURNAL OF DIGESTOLOGY
- Issue:
- 32
- DOI:
- 10.3969/j.issn.1009-3079.2007.32.007
- Key Word:
- 肝癌;细胞周期蛋白E;RNA干扰
Abstract： 目的:以cyclinE基因编码区为靶位,构建表达小干扰RNA(siRNA)的质粒载体,观察转染后对HepG2细胞的影响.方法:针对cyclinE基因序列构建表达siRNA的真核表达载体pSilencer3.1-H1hygro,利用脂质体Metafectene转染cyclinE基因高表达的肝癌细胞株HepG2.流式细胞仪检测细胞周期及凋亡率,MTT法检测细胞增殖活性,RT-PCR和Western blot法观察转染后细胞cyclinE基因表达.结果:成功构建了表达siRNA的真核质粒载体,转染后cyclinE基因mRNA及蛋白表达水平分别下降了79%和65%.结论:靶向cyclinE基因的siRNA可有效沉默HepG2细胞高表达的cyclinE基因,从而抑制肝癌细胞的增殖并促进凋亡.
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