Pan Guangliang, Shawer Mohannad, Oie Svein, Lu D Robert
Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, Georgia 30602, USA.
Pharm Res. 2003 May;20(5):738-44. doi: 10.1023/a:1023477317668.
To develop and evaluate a novel artificial lipoprotein delivery system for in vitro gene transfection in human glioma cells.
Nanoemulsion was formulated with similar lipid compositions present in natural lipoproteins. The oil phase of nanoemulsion was composed of triolein (70%), egg phosphatidylcholine (22.7%), lysophosphatidylcholine (2.3%), cholesterol oleate (3.0%), and cholesterol (2.0%). To replace the surface protein as in natural lipoprotein, poly-L-lysine was modified to add palmitoyl chains at a basic condition and was incorporated onto the nanoemulsion particles through hydrophobic interaction. A model plasmid DNA, pSV-beta-Gal containing a reporter gene for beta-galactosidase was carried by the nanoemulsion/poly-L-lysine particles. The charge variation of soformed complex was examined by agarose gel electrophoresis and zeta potential measurement. In vitro transfection was conducted on human SF-767 glioma cell line using this new system. After standard X-Gal staining, transfected cells were observed under light microscope. The effect of chloroquine on the transfection was examined and, finally, the cytotoxicity of this new system was evaluated in comparison with commercial Lipofectamine gene transfection system.
The plasmid DNA was effectively carried by this artificial lipoprotein delivery system and the reporter gene was expressed in the glioma cells. Transfection efficiency was significantly increased by the treatment of chloroquine, indicating that endocytosis possibly was the major cellular uptake pathway. Compared to Lipofectamine system, this new delivery system demonstrated similar transfection efficiency but a much lower cytotoxicity. In the experiment, the cell viability showed up to 75% using this system compared to only 24% using Lipofectamine system.
A new artificial lipoprotein delivery system was developed for in vitro gene transfection in tumor cells. The new system showed similar transfection efficiency but a much lower cytotoxicity compared with commercial Lipofectamine system.
开发并评估一种用于人胶质瘤细胞体外基因转染的新型人工脂蛋白递送系统。
用天然脂蛋白中存在的相似脂质成分制备纳米乳剂。纳米乳剂的油相由三油酸甘油酯（70%）、鸡蛋卵磷脂（22.7%）、溶血卵磷脂（2.3%）、油酸胆固醇酯（3.0%）和胆固醇（2.0%）组成。为了替代天然脂蛋白中的表面蛋白，聚-L-赖氨酸在碱性条件下进行修饰以添加棕榈酰链，并通过疏水相互作用整合到纳米乳剂颗粒上。携带β-半乳糖苷酶报告基因的模型质粒DNA，pSV-β-Gal，由纳米乳剂/聚-L-赖氨酸颗粒携带。通过琼脂糖凝胶电泳和zeta电位测量检测所形成复合物的电荷变化。使用该新系统对人SF-767胶质瘤细胞系进行体外转染。在标准X-Gal染色后，在光学显微镜下观察转染细胞。检测了氯喹对转染的影响，最后，与商业脂质体基因转染系统相比，评估了该新系统的细胞毒性。
该人工脂蛋白递送系统有效地携带了质粒DNA，并且报告基因在胶质瘤细胞中表达。氯喹处理显著提高了转染效率，表明内吞作用可能是主要的细胞摄取途径。与脂质体系统相比，该新递送系统显示出相似的转染效率，但细胞毒性低得多。在实验中，使用该系统时细胞活力高达75%，而使用脂质体系统时仅为24%。
开发了一种用于肿瘤细胞体外基因转染的新型人工脂蛋白递送系统。与商业脂质体系统相比，新系统显示出相似的转染效率，但细胞毒性低得多。
