Fortunati E, Bout A, Zanta M A, Valerio D, Scarpa M
Department of Pediatrics, CRIBI Biotechnology Center, Padova, Italy.
Biochim Biophys Acta. 1996 Apr 10;1306(1):55-62. doi: 10.1016/0167-4781(95)00217-0.
Cationic liposomes have been proposed as alternative to adenovirus in the treatment of cystic fibrosis lung disease. Therefore, we have investigated the efficiency of two lipid mixtures in mediating gene transfer in in vitro and in vivo models. The cationic lipid DOTMA (N-(1-(2,3(dioleyloxy)propyl)-n,n,n-trimethylammoniumchloride++ +) and DOGS (dioctadecylamidoglycylspermine) were used in combination with the neutral lipid DOPE (dioleoylphosphatidylethanolamine). The relative transfection efficiencies of the two cationic liposomes were tested using the bacterial beta-galactosidase (lacZ) and the firefly luciferase genes. Gene expression was detected in both cell limes and primary culture of rhesus monkey airway epithelium after transfection with plasmid DNA complexed with DOGS/DOPE or DOTMA/DOPE. Transfection efficiency of both types of lipids was higher in the mouse fibroblast 3T3 cell line as compared to human carcinoma A549 cells and primary epithelial cultures. Administration of DNA-liposome complexes via intratracheal instillation resulted in expression of the lacZ and luciferase marker gene in the mouse airways. In vivo transfection mediated by both types of liposomes were proven to be far less efficient than adenovirus treatment.
阳离子脂质体已被提议作为治疗囊性纤维化肺病的腺病毒替代物。因此，我们研究了两种脂质混合物在体外和体内模型中介导基因转移的效率。阳离子脂质DOTMA（N-(1-(2,3-二油酰氧基)丙基)-N,N,N-三甲基氯化铵）和DOGS（二辛基酰胺基甘氨酰精胺）与中性脂质DOPE（二油酰磷脂酰乙醇胺）联合使用。使用细菌β-半乳糖苷酶（lacZ）和萤火虫荧光素酶基因测试了两种阳离子脂质体的相对转染效率。用与DOGS/DOPE或DOTMA/DOPE复合的质粒DNA转染后，在恒河猴气道上皮细胞系和原代培养物中均检测到基因表达。与人类癌A549细胞和原代上皮培养物相比，两种脂质在小鼠成纤维细胞3T3细胞系中的转染效率更高。通过气管内滴注给予DNA-脂质体复合物导致lacZ和荧光素酶标记基因在小鼠气道中表达。事实证明，两种脂质体介导的体内转染效率远低于腺病毒治疗。
