摘要
目的 构建绿色荧光蛋白和瑞典突变型APP695共表达载体 ,将重组质粒转染PC12细胞获得高表达APP的PC12细胞克隆。观察基因转染后PC12细胞的细胞周期、超微结构和 β淀粉样蛋白的分泌情况。 方法 激光共聚焦显微镜挑选高表达绿色荧光蛋白的PC12细胞克隆。放射免疫方法测定 β淀粉样蛋白 ,流式细胞仪检测细胞周期 ,电镜观察细胞超微结构。结果 瑞典突变型APP转染能使PC12细胞体积增大 ,形态变长 ,微绒毛增多 ,分泌Aβ增多 ,细胞周期中G0和G1 期细胞比例增多。结论 绿色荧光蛋白和瑞典突变型APP695共表达载体转染的PC12细胞可作为AD发病机制和治疗研究的细胞模型。
Objective To construct an eukaryotic expression vector carrying human mutant APP695(SW) and enhanced green fluorescence protein(EGFP) and to transfect the PC12 cells. Methods Eukaryotic expression vector was transfected into PC12 cell by liposome mediated method. The APP overexpressed clones were selected by the magnitude of green fluorescence protein under laser scanning confocal microscope. The concentration of Aβ was measured by radioimmunoassay. Ultrastructural changes and cell cycle were observed by electron microscopy and flow cytometry. Results It was found that the transfection of PC12 cells with APP695 gene induced the increased secretion of Aβ and the increase of resembled microvilli morphologically. More proportion of transfected cells was found in G 0 G 1 phase compared with untransfected PC12 cells. Conclusion PC12 cells transfected with human mutant APP695(SW) gene can be used as the cellular model for the studies of the pathogenesis and therapy in Alzheimer disease(AD).
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2003年第3期205-208,共4页
Journal of Third Military Medical University
基金
国家自然科学基金资助项目 ( 3990 0 16 9
30 10 0 0 87)
全军"十五"青年基金资助项目 ( 0 1Q0 99)
重庆市应用基础基金资助项目 ( 2 0 0 1)
