| 环氧合酶-2特异性siRNA真核表达质粒对HT-29细胞生长的影响 |
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| 引用本文： | 王磊,陈卫昌,杨吉成. 环氧合酶-2特异性siRNA真核表达质粒对HT-29细胞生长的影响[J]. 肿瘤防治研究, 2008, 35(10): 705-710. DOI: 10.3971/j.issn.1000-8578.3326 |
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| 作者姓名： | 王磊 陈卫昌 杨吉成 |
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| 作者单位： | 苏州大学附属第一医院消化科; 苏州大学医学院细胞与分子生物学教研室; |
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| 基金项目： | 江苏省卫生厅"135工程"医学重点人才基金 |
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| 摘 要： | 目的探讨环氧合酶-2(COX-2)特异性小干扰RNA(siRNA)真核表达质粒对人结肠癌HT-29细胞COX-2表达和生长的影响。方法设计短发夹结构的COX-2 siRNA对应模版DNA序列,构建重组质粒pshCOX-2。将重组质粒转染人结肠癌HT-29细胞,采用逆转录-聚合酶链反应(RT-PCR)、Western印迹分别从mRNA和蛋白水平检测COX-2表达。用四氮唑盐(MTT)法和流式细胞仪观察COX-2表达被抑制后细胞的生长情况,放射免疫法(RIA)和ELISA法分别检测培养上清中PGE2和VEGF的含量变化。结果酶切及测序证实质粒pshCOX-2构建成功。转染重组质粒72h后可以显著抑制HT-29细胞COX-2 mRNA和蛋白表达(P<0.05),细胞生长受抑,凋亡细胞数显著增加(P<0.05),细胞PGE2和VEGF的产生受抑制(P<0.05)。结论成功构建的COX-2 siRNA真核表达质粒pshCOX-2通过抑制人结肠癌HT-29细胞COX-2基因表达,从而抑制细胞生长、PGE2合成和VEGF的产生。
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| 关 键 词： | 环氧合酶-2 siRNA RNA干扰 真核表达质粒 结肠癌 |
| 收稿时间： | 2007-08-07 |
| 修稿时间： | 2008-06-03 |
Effect of Eukaryotic Expression Plasmid for Cyclooxygenase-2 Specific siRNA on Growth of HT-29 Cells in vitro |
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| WANG Lei,CHEN Wei-chang,YANG Ji-cheng. Effect of Eukaryotic Expression Plasmid for Cyclooxygenase-2 Specific siRNA on Growth of HT-29 Cells in vitro[J]. Cancer Research on Prevention and Treatment, 2008, 35(10): 705-710. DOI: 10.3971/j.issn.1000-8578.3326 |
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| Authors: | WANG Lei CHEN Wei-chang YANG Ji-cheng |
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| Affiliation: | .Department of Gastroenterology; The First Affiliated Hospital of Soochow University; Soochow 215006; China; 2.Department of Cellular and Molecular Biology; Soochow University; |
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| Abstract: | Objective To investigate the effect of the eukaryotic expression plasmid of specific small interfering RNA(siRNA)against COX-2 gene on the COX-2 expression and growth of human colon cancer HT-29 cells.Methods The COX-2 siRNA template DNA sequence for short hairpin RNA(shRNA) was designed and synthesized.The recombinant plasmid(pshCOX-2) was transfected into HT-29 cells.The effect of the recombinant plasmid on the COX-2 expression of human colon cancer HT-29 cells was detected by RT-PCR and Western blot.Chan... |
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| Keywords: | siRNA |
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