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。Lipofectamine 2000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Lipofectamine 2000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Lipofectamine 2000与DOTAP都能有效压缩DNA,形成复合物。Lipofectamine 2000与DOTAP相比,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3/1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,Lipofectamine 2000对Hep-2细胞的转染效率比NCI-H460高。
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.
出处
《中国细胞生物学学报》
CAS
CSCD
北大核心
2013年第10期1459-1464,共6页
Chinese Journal of Cell Biology
基金
国家自然科学基金(批准号:20876027
21176046)
中央高校自主科研基金(批准号:DC12010104)资助的课题~~
