转基因载体聚乙二醇-聚谷氨酸纳米微球的制备  

Study on Encapsulation of Plasmid DNA in Poly(Ethylene Glycol)-Poly(Benzyl L-Glutamate) Copolymer Nanospheres and its Characteristics in Vitro

作　　者：余东升[1] 黄洪章[1] 李苏[2] 王安训[3] 潘朝斌[1] 王建广[1] 潘仕荣[3] 

机构地区：[1]中山大学第二附属医院口腔颌面外科,510120 [2]中山大学肿瘤医院内科实验室,510080 [3]中山大学第一附属医院,510080

出　　处：《实用癌症杂志》2004年第1期5-8,共4页The Practical Journal of Cancer

基　　金：国家自然科学基金 (No .30 2 71 4 2 3);广东省自然科学基金资助 (No .2 1 865)

摘　　要：目的　制备聚乙二醇 聚谷氨酸两嵌段共聚物 (PEG PBLG )纳米微球并观察其转基因能力。方法　合成两亲嵌段共聚物PEG PBLG ,红外光谱 (IR)、核磁共振谱 ( 1H NMR )、凝胶渗透色谱法 (GPC)测定其组成和结构 ;乳化溶剂蒸发法制备DNA/PEG PBLG纳米微球 ,透射电镜观察其形态 ,DNaseⅠ消化实验测试其DNA保护能力 ,以GFP为报告基因转染Tca8113细胞观察其转基因能力。结果　IR图谱证实两嵌段共聚物的形成 ,GPC和1H NMR测定PEG PBLG分子量约 80 0 0 ,PEG PBLG纳米微球直径约 70nm ,对质粒DNA有较好的保护作用并有较强转基因能力。结论　成功制备两亲嵌段共聚物PEG PBLG纳米微球并证实其载基因能力 。Objective To prepare poly(ethylene glycol)-poly(benzyl L-glutamate) amphipathic copolymer nanosphere and evaluate its gene-delivery ability.Methods The amphipathic copolymers were prepared by polymerizing of benzyl L-glutamate N-carboxyanhydride(BLG-NCA) initiated by AT-PEG.The characteristics of the copolymers were measured by IR, 1H-NMR and GPC.The copolymer nanospheres were prepared by the solvent evaporation technique.The microscopic morphology of the nanoparticles were examined by TEM.DNA protection was observed by DNaseⅠdigestion.The GFP gene was used as reporter gene to analyze the gene-delivery ability.Results The copolymers nanospheres are core-shell structures and the diameters are in the range of 60～80 nm.The plasmid DNA can be protected by the PEG-PBLG nanospheres from DNaseⅠ digestion.The GFP expression efficacy was found as well as that of lipofentamine teansfection.Conclusion The PEG-PBLG nanospheres display perfect ability to encapsulate plasmid DNA and can be potentially used as a new vector in gene therapy.

关 键 词：转基因载体聚 乙二醇—聚谷氨酸纳米微球 制备 核磁共振谱 外光谱 恶性肿瘤 基因治疗 

分 类 号：R73-33[医药卫生—肿瘤]