制备肝靶向载体半乳糖基化壳聚糖/DNA纳米颗粒转染肝癌细胞SMMC-7721的体外实验  

PREPARATION OF HEPATOCYTE-TARGETING GENE VECTOR GALACTOSYLATED CHITOSAN/DNA NANOPARTICLES FOR TARGETING SMMC-7721 CELLS IN VITRO

作　　者：徐缨龙[1] 蒋辉[1] 汪景洲[1] 黄纪伟[1] 吴泓[1] 曾勇[1] 

机构地区：[1]四川大学华西医院肝胆胰外科,成都610041

出　　处：《现代预防医学》2010年第18期3507-3510,共4页Modern Preventive Medicine

基　　金：四川省科技厅项目(2008)

摘　　要：[目的]研究半乳糖基化壳聚糖(Galactosylated chitosan,GC)与质粒DNA混合制备纳米颗粒的方法及其对肝癌细胞SMMC-7721转染效率的体外实验。[方法]水溶性GC溶液与质粒pEGFP通过磁力搅拌(1000rpm,室温)并逐滴滴加混合制备为纳米微囊复合物;制备的GC/DNA纳米微囊体外转染肝癌细胞SMMC-7721,以脂质体转等量质粒及裸质粒DNA为对照,荧光显微镜下观察绿色荧光蛋白的表达情况,流式细胞仪测定转染效率。[结果]半乳糖基化壳聚糖与质粒DNA在一定物理作用条件下混合能够形成纳米微囊复合物;SMMC-7721实验组与对照组均有绿色荧光蛋白表达,实验组(GC/DNA纳米微囊组)转染率为(6.60±0.56)%,对照组(脂质体组)转染率为(20.62±2.27)%。[结论]GC/DNA纳米微囊能将质粒DNA转入肝癌细胞细胞内并表达,可作为基因治疗的一个载体。[Objective] To investigate the way to develop nanoparticles by complex GC with plasmid pEGFP,and the transfection efficiency of the GC /DNA nanoparticles in vitro.[Methods] The GC /DNA nanoparticles were prepared by adding pEGFP solution dropwise to water soluble GC solution with magnetic stirring at 1 000 rpm at ambient temperature in different N /P ratio.The newly prepared GC /DNA nanoparticles were transfected into SMMC-7721 cells（GC group）,LipofectaneTM 2 000（Lipofectane group） and naked DNA（Naked DNA group） were used as controls.Green fluorescence was observed under the inverted confocal laser scanning microscopy,and the transfection was efficiency wcalculated by flow cytometry.[Results] GC was mixed with plasmid DNA could form nanoparticles stable under certain physical conditions.Green fluorescent were detected in both GC group（6.60 ± 0.56）% and Lipofectane group（20.62 ± 2.27）%.[Conclusion] GC /DNA nanoparticles could transfect plasmid DNA into hepatocarcinoma cell SMMC-7721,and it would be a potential carrier for gene therapy.

关 键 词：基因治疗 壳聚糖 绿色荧光蛋白 

分 类 号：R735.7[医药卫生—肿瘤]