目的:利用阳离子多聚物多聚乙烯亚胺(PEI)纳米凝胶将增强型绿色荧光蛋白(enhanced green fluorescence protein,EGFP)转入同步化的肿瘤细胞中并检测绿色荧光蛋白的表达率,探讨PEI作为载体介导肿瘤细胞基因转染中细胞周期对转染效率的影...目的:利用阳离子多聚物多聚乙烯亚胺(PEI)纳米凝胶将增强型绿色荧光蛋白(enhanced green fluorescence protein,EGFP)转入同步化的肿瘤细胞中并检测绿色荧光蛋白的表达率,探讨PEI作为载体介导肿瘤细胞基因转染中细胞周期对转染效率的影响。方法:通过6 MV X线照射及药物阻滞使Hela及A549细胞同步化,在5个PEI/DNA梯度比(2/2、4/2、6/2、8/2、10/2μg/μg)下利用PEI介导EGFP报告基因转染进入上述同步化细胞,流式细胞仪和荧光显微镜下测定绿色荧光蛋白表达率,通过梯度试验分析细胞周期时相与表达率之间的关系。结果:在不同增殖状态及PEI/DNA比值梯度下,一定剂量-时间的X射线和顺铂可以使Hela细胞同步化于S期,一定剂量-时间的艾素可以使A549细胞同步化于G2/M期,而血清饥饿法培养则可以使细胞阻滞于G0/G1期,在不同细胞周期时相下EGFP的转染效率不同,两种同步化细胞均表现出S期及G2/M期细胞的转染率显著高于对照组(P<0.05)和G0/G1期细胞(P<0.0001)。结论:PEI导入基因的表达效率受细胞周期制约,PEI介导的基因转染受到宿主细胞摄取、转运入核及转录能力的影响,而该进程都是细胞周期依赖性的。展开更多
Based on the noncovalent functionalization of ferrocene-grafted polyethylenimine （PEI-Fc） and carbon nanotubes （CNTs）, CNT bundles are exfoliated by PEI-Fc solution and thus form stable compounds PEI-Fc@CNTs, whic...Based on the noncovalent functionalization of ferrocene-grafted polyethylenimine （PEI-Fc） and carbon nanotubes （CNTs）, CNT bundles are exfoliated by PEI-Fc solution and thus form stable compounds PEI-Fc@CNTs, which is used to construct the PEI-Fc@CNTs/DNA multilayers through layer-by-layer assembly. The multilayers show a highly uniform and homogeneous characteristic, which significantly improve the electrical property of the multilayers. Upon the oxidation electrical potential, the ferrocene groups are switched from reduction state （[Fe（C5H5）2]） to oxidation state （[Fe（C5H5）2]^＋）, leading to change of microenvironments＇ charge density, resulting in swelling of the multilayers and a final degree of swelling of 37 % and the decrease of multilayer stiffness. We maintain that electrochemical control over the swelling behavior of multilayers could have important implications for responsive coatings of nanoscale devices, including mechanically tunable surfaces which are used to modulate cellular activities and control drug delivery.展开更多
基金supported by the National Natural Science Foundation of China(21174126,51333005,21374095)the National Basic Research Program of China(2011CB606203)+2 种基金Research Fund for the Doctoral Program of Higher Education of China(20110101110037,20120101130013)the Qianjiang Excellence Project of Zhejiang Province(2013R10035)International Science&Technology Cooperation Program of China(2014DFG52320)
文摘Based on the noncovalent functionalization of ferrocene-grafted polyethylenimine （PEI-Fc） and carbon nanotubes （CNTs）, CNT bundles are exfoliated by PEI-Fc solution and thus form stable compounds PEI-Fc@CNTs, which is used to construct the PEI-Fc@CNTs/DNA multilayers through layer-by-layer assembly. The multilayers show a highly uniform and homogeneous characteristic, which significantly improve the electrical property of the multilayers. Upon the oxidation electrical potential, the ferrocene groups are switched from reduction state （[Fe（C5H5）2]） to oxidation state （[Fe（C5H5）2]^＋）, leading to change of microenvironments＇ charge density, resulting in swelling of the multilayers and a final degree of swelling of 37 % and the decrease of multilayer stiffness. We maintain that electrochemical control over the swelling behavior of multilayers could have important implications for responsive coatings of nanoscale devices, including mechanically tunable surfaces which are used to modulate cellular activities and control drug delivery.
