基础研究(Basic Research)

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

Published at: 2015年第24卷第8期

杨蔚 1,2 , 张毅 3 , 钟玲 4 , 吴边 2 , 杨金伟 2 , 赵四晖 2 , 陈嘉勇 3
1 昆明医科大学,云南 昆明 650500
2 云南省第一人民医院普外二科,云南 昆明 650032
3 昆明医科大学第二附属医院 急诊科,云南 昆明 650101
4 云南省第一人民医院急诊ICU,云南 昆明 650032
通讯作者 嘉勇 陈 Email: 24795812@qq.com
DOI: 10.3978/j.issn.1005-6947.10.3978/j.issn.1005-6947.2015.08.012
基金:
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

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WTXmRNA 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。


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

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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引用

引用本文: 蔚 杨, 毅 张, 玲 钟, 边 吴, 金伟 杨, 四晖 赵, 嘉勇 陈. 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.
Cite this article as: YANG Wei, ZHANG Yi, ZHONG Ling, WU Bian, YANG Jinwei, ZHAO Sihui, CHEN Jiayong . Comparison of electroporation and chemical agent transfection in gastric cancer cells with multidrug resistance[J]. Chin J Gen Surg, 2015, 24(8): 1116-1120.