PEI-CS/siRNA复合颗粒对肝癌耐药细胞BEL7402/5-FU中MRE11表达的影响

目的:探讨聚乙烯亚胺-壳聚糖(PEI-CS)/si RNA复合颗粒对肝癌耐药细胞BEL7402/5-FU中MRE11表达的影响。方法:采用复凝聚法将PEI-CS(100μg/m L)与不同浓度的MRE11 si RNA-FAM形成PEI-CS/si RNA复合颗粒,并转染BEL7402/5-FU细胞,用荧光显微镜和Real-time PCR检测转染效率和沉默效率。结果:荧光显微镜观察结果显示:转染细胞48 h后,3.125、6.25、12.5、25、50μg/m L的si RNA与PEI-CS形成的复合颗粒的转染率分别为62.31%、76.09%、79.99%、86.49%、96.59%。转染细胞48、72、96 h后,12.5μg/m L的si RNA与PEI-CS形成的复合颗粒的转染率分别为78.22%、55.76%、42.85%,25μg/m L的si RNA与PEI-CS形成的复合颗粒的转染率分别为83.67%、74.23%、67.45%。Real-time PCR检测结果显示:25μg/m L的si RNA与PEI-CS形成的复合颗粒转染48小时后,对BEL7402/5-FU细胞中MRE11基因的沉默效率为35.4%。结论:聚乙烯亚胺-壳聚糖/si RAN复合颗粒能有效转染肝癌耐药细胞Bel7402/5-FU,并对BEL7402/5-FU细胞中MRE11基因表达有一定抑制作用。

Effect of PEI-CS/siRNA Composite Particles on Multidrug-resistant Hepatocellular Carcinoma Cell Line BEL7402/5-FU in MRE11 Gene Expression

Objective:To explore the effect of PEI-CS/siRNA nanoparticles on gene expression of multidrug-resistant hepatocellular carcinoma cell line BEL7402/5-FU in MRE11.Methods:BEL7402/5-FU was transfected with PEI-CS/siRNA nanoparticles, which were prepared with PEI-CS (100 ug/mL) and different concentration of MRE11 siRNA-FAM by condensation method. Then, the rate of transfection was observed by fluorescence microscopy, the silencing efficiency was assessed by Real-time PCR.Results:As shown by fluorescence microscopy, when the cell was transfected after 48 hours using nanoparticles which were prepared with PEI-CS at different concentration of siRNA (3.125, 6.25, 12.5, 25, 50 ug/mL), the transfection efficiency showed 62.31%, 76.09%, 79.99%, 86.49%, and 96.59%, respectively. When the cell was transfected with nanoparticles at the concentration of 12.5 ug/mL for different times (48, 72, 96 h), the transfection efficiency was 78.22%, 55.76% and 42.85%, respectively. However, under the test conditions above, the transfection efficiency were respectively turned into 83.67%, 74.23% and 67.45%, when the concentration of nanoparticles was 25 ug/mL. Real-time PCR results showed that the efficiency of RNA interference for MRE11 was 35.4% by PEI-CS/siRNA nanoparticles at the concentration of 25 ug/mL.Conclusion:PEI-CS/siRNA nanoparticles can not only effectively transfectmultidrug-resistant hepatocellular carcinoma cell line BEL7402/5-FU, but also down-regulate expression levels of geneMRE11.