| 靶向cyclinE小干扰RNA对肝癌HepG2细胞增殖和凋亡的影响 |
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| 引用本文： | 田雪飞,黄燕,朱才,肖竺,郭永良. 靶向cyclinE小干扰RNA对肝癌HepG2细胞增殖和凋亡的影响[J]. 世界华人消化杂志, 2007, 15(32): 3385-3389 |
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| 作者姓名： | 田雪飞 黄燕 朱才 肖竺 郭永良 |
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| 作者单位： | 1. 湖南中医药大学中西医结合学院,湖南省长沙市,410208
2. 中南大学湘雅医院感染科,湖南省长沙市,410008 |
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| 基金项目： | 湖南省自然科学基金 , 湖南省教育厅青年基金 |
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| 摘 要： | 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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| 关 键 词： | 肝癌 细胞周期蛋白E RNA干扰 |
| 收稿时间： | 2007-07-20 |
| 修稿时间： | 2007-10-29 |
Effects of small RNA interference to target cyclinE gene on apoptosis and proliferation of HepG2 hepatocellular carcinoma cells |
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| Xue-Fei Tian,Yan Huang,Cai Zhu,Zhu Xiao,Yong-Liang Guo. Effects of small RNA interference to target cyclinE gene on apoptosis and proliferation of HepG2 hepatocellular carcinoma cells[J]. World Chinese Journal of Digestology, 2007, 15(32): 3385-3389 |
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| Authors: | Xue-Fei Tian Yan Huang Cai Zhu Zhu Xiao Yong-Liang Guo |
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| Abstract: | AIM:To design pSilencer3.1-H1hygro plasmid expressing small interfering RNA(siRNA)that targets the coding region of cyclinE,and to evaluate the inhibitory effect of this siRNA on HepG2 cells after transfection. METHODS:pSilencer3.1-Hlhygro vector of the siRNA based on the coding region of cyclinE was constructed.The plasmid and liposome metafectene were cotransfected into HepG2 cells,which overexpressed cyclinE.Cell prolif- eration and apoptosis were assessed by MTT assay and flow cytometry,respectively,and ex- pression of cyclinE was detected by RT-PCR and Western blotting. RESULTS:The plasmid expressing siRNA targeted on the coding region of cyclinE was successfully constructed.Expression of cyclinE mRNA and protein was suppressed by up to 79% and 65%,respectively. CONCLUSION:siRNA targeted on cyclinE can silence cyclinE overexpression,and subsequent- ly suppress proliferation and promote apoptosis of HepG2 cells. |
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| Keywords: | Hepatocellular carcinoma cyclinE RNA interference |
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