绿色荧光蛋白报道基因在鼠C6胶质瘤细胞中的应用研究

Study on the Application of GFP Reporter Gene as a Tag for Rat C6 Glioma Cells

目的 :探讨绿色荧光蛋白 (GFP)报道基因转染C6鼠胶质瘤细胞的体内外表达及对细胞生物学性状的影响。方法 :荧光相差显微镜筛选稳定表达GFP的克隆 ,流式细胞术分析细胞周期。将已转染和未转染瘤细胞植入SD大鼠脑内 ,建立动物模型 ,定期随机处死大鼠 ,鼠脑标本行病理学、增殖与凋亡的检测 ,激光共聚焦显微镜检查肿瘤细胞内荧光强度及其分布。结果 :GFP在C6瘤细胞的体外和体内均获得长期稳定表达 ,检测肿瘤细胞的敏感性及特异性优于HE染色。结论

Aim:To investigate the expression of GFP reporter genes transfected in rat C6 glioma cells as a tag and its effect on biological characteristics of tumor cells.Methods:Transfected clone with strong GFP expression was selected.Cell cycle was analyzed by flowcytometry.GFP-transfected cells and parental cells were implanted into SD rat brains to establish glioma models.The rats were killed randomly at regular interval.Tumor samples were examined for patholgical changes,proliferation activity and apoptosis.Confocal laser scanning microscope was used for detection the distribution of tagged cells and its fluorescence intensity.Results:GFP-trancfected C6 glioma cells maintained a long term high-level GFP expression in vivo and in vitro.It's easier to show the primary tumor margins and the distant micrometastasis and local invasion,its sensitivity and specificity for examination of tumor cells is superior to HE staining.Conclusion:GFP gene is an excellent reporter gene without effect on the biological characteristics of tumor cells.