| E2F-1基因沉默对舌鳞癌细胞环氧化酶-2基因表达影响的研究 |
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| 引用本文： | 杜一飞,周薇娜,江飞,袁华.E2F-1基因沉默对舌鳞癌细胞环氧化酶-2基因表达影响的研究[J].口腔医学,2011,31(10):588-591. |
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| 作者姓名： | 杜一飞 周薇娜 江飞 袁华 |
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| 作者单位： | 1. 南京医科大学口腔医学研究所,南京医科大学口腔医学院口腔颌面外科,南京,210029
2. 南京医科大学口腔医学院口腔修复科,南京,210029 |
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| 基金项目： | 南京医科大学科技发展基金面上项目(09NJMUM121) |
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| 摘 要： | 目的探讨核转录因子E2F-1基因沉默对舌鳞癌细胞环氧化酶-2(COX-2)基因表达的影响。方法体外合成靶向E2F-1基因的短发卡状RNA(short hairpin RNA,shRNA)序列,克隆到pLKO.1-PURO-U6(Age I/EcoR I)质粒载体中,经酶切和测序鉴定所构建的重组载体,将以上质粒分别和包装质粒混合物共转染293T细胞,包装产生病毒颗粒。将病毒载体转染舌鳞癌细胞Tca8113细胞后,运用Real-Time PCR和Western-blot检测E2F-1和COX-2基因的表达。结果经酶切和测序证明,靶向E2F-1基因的shRNA序列正确;转染组E2F-1和COX-2基因mRNA和蛋白表达均发生明显下调,与空白对照组相比,差异有统计学意义,阴性对照组未见明显差异。结论靶向E2F-1基因的shRNA慢病毒载体可特异性沉默E2F-1基因的表达;E2F-1基因沉默后可下调COX-2基因表达。
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| 关 键 词： | E2F-1 shRNA 环氧化酶-2 |
Influence of RNA interference lentiviral vector in E2F-1 on COX-2 in Tca8113 cells |
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| DU Yi-fei,ZHOU Wei-na,JIANG Fei,YUAN Hua.Influence of RNA interference lentiviral vector in E2F-1 on COX-2 in Tca8113 cells[J].Stomatology,2011,31(10):588-591. |
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| Authors: | DU Yi-fei ZHOU Wei-na JIANG Fei YUAN Hua |
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| Institution: | DU Yi-fei,ZHOU Wei-na,JIANG Fei,YUAN Hua.(The Research Institute of Stomatology,Department of Oral and Maxillofacial Surgery,School of Stomatology,Nanjing Medical University,Nanjing 210029,China) |
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| Abstract: | Objective To investigate the influence of RNA interference lentiviral vector in E2F-1 on COX-2 in Tca8113 cells.Methods DNA oligonucleotides targeting gene E2F-1 on appropriate site and one pair of negative control oligonucleotide sequence were synthesized and inserted into pLKO.1-PURO-SP6(Age I/EcoR I) vector.The sequences of two plasmids were identified by DNA sequencer and restriction endonuclease digestion.These recombinant plasmids were transfected into the 293T cells along with lentiviral packing mix ... |
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| Keywords: | E2F-1 shRNA COX-2 |
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