shRNA靶向抑制COX-2基因表达对胃癌细胞SGC-7901增殖的影响
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丘敏梅E-mail:qiumm988@126.com

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Effects of COX-2 gene expression inhibited by shRNA on proliferation of gastric carcinoma SGC-7901 cells
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    摘要:

    目的:探讨环氧化酶-2(COX-2)短发夹RNA(shRNA)对胃癌细胞SGC-7901的生长和细胞周期的影响。
    方法:构建COX-2基因的特异性小RNA干扰质粒,构建靶向抑制COX-2基因的重组表达载体pshRNA-COX-2。实验分3组:未转染胃癌细胞组,阴性对照HK组,pshRNA-COX-2转染组。用质脂体lipofectamineTM2000转染胃癌细胞SGC-7901,RT-PCR和Western blot分别检测COX-2基因mRNA和蛋白表达情况,流式细胞术检测细胞周期,细胞计数检测细胞的生长变化。
    结果:与阴性对照组相比,pshRNA-COX-2重组表达载体抑制COX-2 mRNA及蛋白表达的抑制率分别为70.1%和43.2%;G0~G1期细胞由61.5%上升至70.2%,S期细胞由27.3%下降至21.7%;细胞生长明显减慢。
    结论:pshRNA-COX-2重组表达载体能显著抑制COX-2的表达,从而抑制胃癌细胞生长。

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    Objective:To investigate the inhibiting effects of shRNA (short hairpin RNA, shRNA) on COX-2 gene expression, the cell cycle and growth of gastric carcinoma SGC-7901 cells.
    Methods:Specific shRNA plasmid to COX-2 were constructed, and then transfected into SGC-7901 cells by lipofectamine methods. Tests were divided into three groups: untransfected gastric carcinoma SGC-7901 cells group, negative control HK group and pshRNA-COX-2 group. Gastric carcinoma SGC-7901 cells were transfected with LipofectamineTM 2000. Expression of COX-2 mRNA and protein were detected with reverse transcriptional polymerase chain reaction (RT-PCR) and Western blot respectively. Cell cycle analysis and cell growth chart were detected with flow cytometry and cell count respectively.
    Results:Compared with negative control HK group, recombinant expression vector pshRNA-COX-2 resulted in the reduction of COX-2 mRNA and protein expresion by 70.1% and 43.2% respectively; cells in G0-G1 phase  increased from 61.5%to 70.2%, cells in S phase decreased from 27.3% to 21.7%, and the growth of SGC-7901 cells cells was slowed significantly.
    Conclusions:Recombinant expression vector  pshRNA-COX-2 can significantly inhibit the expression of COX-2 gene, result in the  increase of cells in G0-G1 phase and decrease of cells in S phase, and suppress proliferation of gastric carcinoma SGC-7901 cells.

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丘敏梅, 向华国, 陈亚芹, 杨晓丽. shRNA靶向抑制COX-2基因表达对胃癌细胞SGC-7901增殖的影响[J].中国普通外科杂志,2009,18(12):1280-1283.
DOI:10.7659/j. issn.1005-6947.2009.12.013

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  • 收稿日期:2008-12-11
  • 最后修改日期:2009-04-22
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  • 在线发布日期: 2009-12-15