自组装修饰在不同细胞系中对聚酰胺-胺介导的基因递送的影响  

Effects of self-assembling modifications on PAMAM-mediated gene delivery in different cell lines

作　　者：殷哲[1] 郝艳丽[1] 刘楠[1] 王兰[1] 马茗舒[1] 闫冬[1] 张小宁[1] 

机构地区：[1]清华大学医学院药剂学实验室,北京100084

出　　处：《中国新药杂志》2012年第23期2740-2743,2752,共5页Chinese Journal of New Drugs

基　　金：国家"重大新药创制"科技重大专项(2012ZX09103301-31);国家自然科学基金(31271070)

摘　　要：目的:通过自组装方法使用蛋白质分子修饰聚酰胺-胺树状大分子(PAMAM)与DNA形成的转染复合物,并考察其性质。方法:使用人血白蛋白(HSA)和表皮生长因子(EGF)修饰PAMAM-DNA转染复合物,测定复合物粒径及Zeta电位;通过DNA固缩程度测定试验和DNaseI消化试验考察修饰后复合物的稳定性;HEK 293T细胞与MCF-7细胞转染质粒,测定其荧光素酶表达水平;MTT检测修饰后复合物对细胞毒性的变化。结果:自组装修饰后的复合物的粒径无显著变化,Zeta电位显著降低,性质稳定;HSA修饰可显著提高复合物在HEK 293T和MCF-7细胞中的转染效率,EGF修饰仅显著提高复合物在MCF-7细胞中的转染效率;两种修饰都能降低PAMAM-DNA复合物对细胞的毒性。结论:自组装修饰方法快速、简便、有效,不影响PAMAM-DNA复合物的稳定性,并且能够实现提高转染效率、靶向递送、降低毒性等目的。Objective： To modify PAMAM-DNA transfection complex with protein molecules by self-assem- bly, and to evaluate its property. Methods： PAMAM-DNA transfection complex was modified using human serum albumin （HSA） or epidermal growth factor （EGF） as modification proteins. Size and Zeta-potential of such com- plex was characterized by laser particle analyzer, and stability was tested by DNA condensation and DNaseI diges- tion. The luciferase expression was measured after transfecting luciferase expressing plasmid in HEK 293T and MCF-7 cells. The cytotoxicity of modified complexes was measured by MTT assay. Results： Both HSA and EGF modification exhibited similar sizes, significantly lower Zeta-potentials and equal stability. HSA modification promo- ted transfection in both HEK 293T and MCF-7 cell lines, while EGF modification showed evident transfection en- hancing effect only in MCF-7. Modification with either molecule, HSA or EGF, reduced cytotoxicity of PAMAM-. DNA complex. Conclusion： Self-assembly is a rapid and effective way to modify PAMAM-DNA complexes with protein molecules like HSA or EGF, without sacrificing stability, and successfully reaches the goals of enhancing transfection efficiency, targeted delivery, and lowering cytotoxicity.

关 键 词：聚酰胺-胺 自组装 稳定性 转染效率 细胞毒性 

分 类 号：R945[医药卫生—微生物与生化药学]