文章摘要
潘杰 朱欣婷 刘云 谢星星 侯玉群 李长福 范芳.PEI-CS/siRNA复合颗粒对肝癌耐药细胞BEL7402/5-FU中MRE11表达的影响[J].,2015,15(10):1844-1847
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/siRNA Composite Particles on Multidrug-resistantHepatocellular Carcinoma Cell Line BEL7402/5-FU in MRE11 GeneExpression
  
DOI:
中文关键词: 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
英文关键词: Polyethyleneimine-chitosan  Composite particles  MRE11  RNA interference  BEL7402/5-FU
基金项目: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)
作者单位
潘杰 朱欣婷 刘云 谢星星 侯玉群 李长福 范芳 遵义医学院基础医学院贵州省普通高等学校特色药物肿瘤防治特色重点实验室 遵义医学院医学与生物学研究中心 
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中文摘要:
      目的:探讨聚乙烯亚胺- 壳聚糖(PEI-CS)/siRNA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(100 ug/mL)与不同浓度的MRE11 siRNA-FAM形成PEI-CS/siRNA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、50 ug/mL的siRNA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,12.5 ug/mL 的siRNA 与PEI-CS 形成的复合颗粒的转染率分别为78.22%、55.76%、42.85%,25 ug/mL 的siRNA 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:25 ug/mL 的siRNA 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/siRAN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。
英文摘要:
      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.
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