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阳离子脂质体介导基因转染肿瘤细胞
引用本文:赵轶男,张树彪,崔韶晖,张传敏,张淑芬.阳离子脂质体介导基因转染肿瘤细胞[J].细胞生物学杂志,2013(10):1459-1464.
作者姓名:赵轶男  张树彪  崔韶晖  张传敏  张淑芬
作者单位:[1]大连民族学院生物技术与资源利用国家民委一教育部重点实验室,大连116600 [2]大连理工大学精细化工国家重点实验室,大连116012
基金项目:国家自然科学基金(批准号:20876027、21176046)和中央高校自主科研基金(批准号:Dcl2010104)资助的课题
摘    要:使用基因转运载体运载肿瘤细胞进行转染是基因治疗的关键环节之一。Lipo-fectamine2000和DOTAP作为商品转染试剂,具有较高的转染效率。为了进一步发掘其作为基因转运载体的应用潜力,该文研究了Lipofectamine2000和DOTAP的粒径、Zeta电位及形态,并分别与绿色荧光蛋白基因(pGFP—N2)、荧光素酶基因(pGL3)结合,形成脂质体/DNA复合物,通过载入人喉癌细胞(Hep-2)和人肺癌细胞(NCI—H460),考察了其转染效率和细胞毒性。结果表明,脂质体Lipofectamine2000与DOTAP都能有效压缩DNA,形成复合物。Lipofectamine2000与DOTAP井目比,转染效率高,与DNA最佳转染比例范围为2:1~4:1。毒性实验显示,在N/P大于3/l时,Lipofectamine2000与DOTAP对癌细胞具有一定的细胞毒性。细胞种类对脂质体的转染效率有很大影响,Lipo—fectamine2000对Hep-2细胞的转染效率比NcI—H460高。

关 键 词:阳离子脂质体  脂质体  DNA复合物  基因转运  细胞毒性

Transfection of Genes into Cancer Cells Mediated by Cationic Liposomes
Zhao Yinan,',Zhang Shubiao,*,Cui Shaohui,Zhang Chuanmin,Zhang Shufen.Transfection of Genes into Cancer Cells Mediated by Cationic Liposomes[J].Chinese Journal of Cell Biology,2013(10):1459-1464.
Authors:Zhao Yinan    Zhang Shubiao    Cui Shaohui  Zhang Chuanmin  Zhang Shufen
Institution:1 Key Laboratory of Biotechnology and Bioresources Utilization, The State Ethnic Affairs Commission-Ministry of Education, Dalian Nationalities University, Dalian 116600, China; 2State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116012, China)
Abstract:The transfection of gene vectors into cancer cells is a key step for gene therapy. Lipofectamine 2000 and DOTAP are two commonly used transfection reagents as they have relatively higher transfection efficiency. The size, zeta potential and morphology of Lipofectamine 2000 and DOTAP were researched. Lipoplexes formed by the combination of cationic liposomes with pGFP-N2 were evaluated in terms of transfection efficiency and cytotox- icity by using two types of cells (Hep-2 and NCI-H460). The results indicated that both of Lipofectamine 2000 and DOTAP could condense DNA to form lipoplexes. Lipofectamine 2000 showed higher transfection efficiency com- pared to DOTAP with an optimal mass ratio range of 2/1 to 4/1. Lipofectamine 2000 and DOTAP showed cytotoxici- ty to some degree under the optimal transfection conditions. Cell type was important influence factor for transfection, and Hep-2 cell was easier to be transfected than NCI-H460 with Lipofectamine 2000.
Keywords:cationic liposomes  liposomes/DNA lipoplex  gene transfection  cytotoxicity
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