论文部分内容阅读
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(e GFP)和目的基因WTX m RNA的表达情况。结果:e GFP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 m RNA表达量明显高于两种化学法转染的SGC7901/VCR细胞中的WTX m RNA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。
Objective: To compare the transfection efficiency of electroporation and chemotherapy in multidrug resistance (MDR) gastric cancer cells. Methods: Plasmids expressing human tumor suppressor gene WTX were transfected into gastric cancer cell line SGC7901 / VCR by electroporation and Lipofectamine 2000 respectively, and the expression of enhanced green fluorescent protein (e GFP) and Expression of target gene WTX m RNA. Results: The results of e GFP analysis showed that the transfection efficiency of SGC7901 / VCR cells was significantly lower than that of SGC7901 cells without drug resistance (all P <0.05), but not by electroporation (P> 0.05), and was significantly higher than that of the two chemical transfection methods (all P <0.05) on SGC7901 / VCR cells. The results of RT-PCR showed that the expression of WTX m RNA in SGC7901 / VCR transfected cells was significantly higher than that in SGC7901 / VCR transfected cells (both P <0.05). Conclusion: For MDR gastric cancer cells, electroporation transfection method is not easy to be affected by the cell membrane components, can maintain a good transfection efficiency.