GFP报道基因转染大鼠C6胶质瘤细胞系动态表达的研究

Dynamic Study on C6 Glioblastoma Cells Transfected by Green Fluorescent Protein

目的:研究绿色荧光蛋白报道基因体外转染C6胶质瘤细胞系的动态表达。方法:脂质体转染后荧光显微镜筛选荧光克隆、激光共聚焦显微镜观察荧光蛋白在细胞内分布,并分析荧光蛋白转染细胞后对其增殖能力的影响。结果:荧光蛋白主要分布在细胞核周围区域。冻存后复苏对细胞存活和荧光强度没有明显影响。C6细胞的增殖没有明显变化。结论:荧光蛋白可以作为胶质瘤基因转染时的优选报道基因。

Aim:To study the dynamic expression of green fluorescent protein (GFP) and enhanced green fluorescent protein (EGFP) on C6 glioblastoma cells in vivo. Methods: After transfection by lipofectamine, dynamic expression of fluorescent cells and positive clones were monitored by phase-contrast fluorescent microscopy, and distribution of GFP within the cells were defined by laser scanning confocal microscopy (LSCM). Cell proliferation were also evaluated. Results: LSCM study indicated strong fluorescent intensity distributed at perinuclear region. The positive cells kept fluroscent intensity after thawing from the frozen cells storing in liquid nitrogen for six months. GFP and EGFP transfection seemed have no effect on C6 cell proliferation. Conclusion: GFP and EGFP are excellent candidates as reproter gene for transfer in glioma.