论文部分内容阅读
[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] The research aimed to study the method for preparing nanoparticles by mixing galactosylated chitosan (GC) with plasmid DNA and its in vitro experiment on transfection efficiency of SMMC-7721 cells. [Method] The water-soluble GC solution and plasmid pEGFP were prepared by mixing magnetic nanoparticles (1000rpm, room temperature) and mixed dropwise. The prepared GC / DNA nanocapsules were transfected into SMMC-7721 hepatoma cells in vitro Transfection of liposome with the same amount of plasmid and naked plasmid DNA as a control, under the fluorescence microscope to observe the expression of green fluorescent protein, transfection efficiency was measured by flow cytometry. [Result] The results showed that the hybridization between galactosylated chitosan and plasmid DNA could form nano-microencapsulated complex under certain physical conditions. The expression of green fluorescent protein in SMMC-7721 experimental group and control group was significantly higher than that in experimental group (GC / (6.60 ± 0.56)% in the microcapsule group, and (20.62 ± 2.27)% in the control group (liposome group). [Conclusion] The GC / DNA nanocapsules can transfer plasmid DNA into hepatoma cells and express them, which can be used as a vector for gene therapy.