电穿孔法和化学法在多药耐药胃癌细胞转染中的效果比较
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陈嘉勇, Email: 24795812@qq.com

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云南省科技厅-昆明医科大学联合专项基金资助项目(2010CD165);云南省卫生科技计划资助项目(2012NS046);云南省应用基础研究重点资助项目(2011FA029);云南省应用基础研究面上项目(2013FZ295)。


Comparison of electroporation and chemical agent transfection in gastric cancer cells with multidrug resistance
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    摘要:

    目的:比较电穿孔法和化学法在多药耐药(MDR)胃癌细胞中转染效果。 方法:将表达抑癌基因WTX的质粒分别用电穿孔法和两种化学法(Lipofectamine2000、Attractene)转染MDR胃癌SGC7901/VCR细胞,观察转染后细胞中增强型绿色荧光蛋白(eGFP)和目的基因WTX mRNA的表达情况。 结果:eGFP分析结果显示,与无耐药的亲代SGC7901细胞比较,两种化学转染法对SGC7901/VCR细胞的转染效率明显降低(均P<0.05),而电穿孔转染法未见明显降低(P>0.05),且明显高于两种化学转染法对SGC7901/VCR细胞的转染效率(均P<0.05)。RT-PCR结果显示,电穿孔转染的SGC7901/VCR细胞中WTX mRNA表达量明显高于两种化学法转染的SGC7901/VCR细胞中的WTX mRNA表达量(均P<0.05)。 结论:对于MDR胃癌细胞,电穿孔转染法不易受其细胞膜成分的影响,可以保持较好的转染效率。

    Abstract:

    Objective: To compare the transfection efficiency of electroporation and chemical transfection agents in multidrug-resistant (MDR) gastric cancer cells.
    Methods: MDR gastric cancer SGC7901/VCR cells were transfected with plasmids bearing tumor suppressor gene WTX through electroporation method and two chemical agents (Lipofectamine2000 and Attractene), respectively. Then, the expression of enhanced green fluorescent protein (eGFP) and the WTX mRNA in the cells after transfection were examined.
    Results: Results of eGFP analysis showed that in SGC7901/VCR cells compared with the parental non-drug-resistant SGC7901 cells, the transection efficiencies of both chemical methods were significantly decreased (both P<0.05), but the transfection efficiency of electroporation exhibited no significant decrease (P>0.05), which was significantly higher than that of either chemical agent (both P<0.05). Results of RT-PCR showed that the WTX mRNA expression level in SGC7901/VCR cells undergoing electroporation transfection was significantly higher than that in those transfected by the two chemical agents (both P<0.05).
    Conclusion: In MDR gastric cancer cells, electroporation transfection will not be influenced easily by their cell membrane components, and can maintain its transfection efficiency.

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杨蔚,张毅,钟玲,吴边,杨金伟,赵四晖,陈嘉勇.电穿孔法和化学法在多药耐药胃癌细胞转染中的效果比较[J].中国普通外科杂志,2015,24(8):1116-1120.
DOI:10.3978/j. issn.1005-6947.2015.08.012

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  • 收稿日期:2015-06-03
  • 最后修改日期:2015-07-17
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  • 在线发布日期: 2015-08-15