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

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

作　　者：潘杰[1] 朱欣婷[1,2] 刘云[2,3] 谢星星[3] 侯玉群 李长福[1,2] 范芳[1,2] 

机构地区：[1]遵义医学院基础医学院,贵州遵义563099 [2]贵州省普通高等学校特色药物肿瘤防治特色重点实验室,贵州遵义563099 [3]遵义医学院医学与生物学研究中心,贵州遵义563099

出　　处：《现代生物医学进展》2015年第10期1844-1847,共4页Progress in Modern Biomedicine

基　　金：贵州省科技厅社会发展攻关项目(黔科合SY[2013]3008);遵义医学院招标项目(F-614);遵义市红花岗区科技基金(遵红科合社字[2009]18);贵州省教育厅特色重点实验室建设项目(黔教合KY字[2014]212)

摘　　要：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。Objective： To explore the effect of PEI-CS/si RNA 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/si RNA nanoparticles, which were prepared with PEI-CS（100 μg/m L） and different concentration of MRE11 si RNA-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 si RNA（3.125, 6.25, 12.5, 25, 50 μg/m L）, 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 μg/m L 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 μg/m L. Real-time PCR results showed that the efficiency of RNA interference for MRE11 was 35.4% by PEI-CS/si RNA nanoparticles at the concentration of 25 μg/m L. Conclusions： PEI-CS/si RNA nanoparticles can not only effectively transfect multidrug-resistant hepatocellular carcinoma cell line BEL7402/5-FU, but also down-regulate expression levels of gene MRE11.

关 键 词：聚乙烯亚胺-壳聚糖 复合颗粒 减数分裂重组蛋白11 RNA干扰 Bel7402/5-FU 

分 类 号：R735.7[医药卫生—肿瘤]