摘要
基因治疗作为一种革命性的治疗手段,对免疫缺陷病、肿瘤等多种疾病的疗效已得到验证。而基因治疗若要大规模进入临床治疗,还需进行很多基础性的研究。其中高效、稳定、无毒、具有靶向性的基因导入载体的研究是基因治疗基础研究进一步深入的关键。基因导入载体主要分为病毒型和非病毒型两种。目前大量应用的病毒类载体的缺陷是免疫原性高、容量小、制备困难、成本较高,而非病毒类基因载体可以克服这些缺点,作为非病毒类基因载体中重要的一种——阳离子脂质体因其操作简便,转染的安全性、高效性和通用性而得到广泛应用。
In order to study the important factors involved in cationic liposome-mediated gene transfer, Lipofectamine 2000 or DOTAP was evaluated using three types of cells (Hep-2, MCF-7 and SW-480) in vitro transfection efficiencies. Different properties of the two reagents were analyzed and compared by DNA arrearage assay and MTT assay. Both Lipofectamine 2000 and DOTAP had strong capability to combine with DNA; Lipofectamine 2000 can get higher transfection efficiency of the three cells by using GFP as report gene, meanwhile, DOTAP can also get higher transfection efficiency against Hep-2 cell. However, DOTAP showed lower transfection efficiency against MCF-7 and SW-480 cell. On the other hand, the cytotoxicity assay showed that over 85% cell viability of MCF-7 cell could be achieved both by Lipofectamine 2000 and DOTAP under the optimal transfection condition. Relatively speaking, Lipofectamine 2000 has very high transfection efficiency in a broad range of cell lines, but because of the special selectivity of cell type on liposome, DOTAP also has a broad application prospect.
出处
《药学学报》
CAS
CSCD
北大核心
2009年第5期553-557,共5页
Acta Pharmaceutica Sinica
基金
国家自然科学基金资助项目(20876027)