Objective To establish a stable and real-time monitorable nude mouse model of orthotopic glioma xenograft.Methods U251 glioma cell line was infected by a lentiviral vector containing green fluorescent protein (GFP) and luciferase (Luc) gene.Cells stably expressing fluorescence of GFP and Luc were sorted by flow cytometry.CCK-8 test and Transwell tumor invasion and migration assay were used to compare the biological features between the cells stably expressing GFP-Luc fluorescence and cells without fluorescence.Then the cells were implanted intracranially in the right caudate nucleus of athymic Balb/c nude mice to establish the tumor model.The growth of intracerebral tumor was monitored over time by a bioluminescence imaging (BLI) system.Hematoxylin-eosin (HE) staining was used to evaluate the histopathological features and tumorigenicity of the transplanted glioma cells in the brain of nude mice.Results U251 glioma cell line with stably expressing GFP-Luc fluorescence and the corresponding orthotopic xenograft model were successfully established.There was no statistically significant difference in the proliferation,invasion and migration abilities between the cells with stably expressing GFP-Luc fluorescence and the control cells.This model showed a high tumor formation rate and stable tumor growth,and takes a moderate time to establish this model.Conclusions Compared with the traditional glioma cells,GFP-Luc-transfected human glioma cells are more feasible for the studies of glioma in vivo.The tumor growth and pathological characteristics in this U251-GFP-Luc glioma model are similar to human glioma,and the growth of this tumor can be real-time monitored.It can be used as an ideal animal model for experimental studies of glioma.%目的 建立一种稳定、可实时监测的胶质瘤原位移植瘤裸鼠模型.方法 用带有荧光素酶(luciferase-Luc)和绿色荧光蛋白(green fluorescent protein-GFP)基因的慢病毒感染U251神经胶质瘤细胞,流式细胞仪筛选稳定表达GFP-Luc荧光的细胞系,并通过CCK-8实验、细胞周期实验、Transwell肿瘤迁移及侵袭实验等评价荧光细胞的增殖、迁移和侵袭能力是否改变;将细胞接种至裸鼠大脑尾状核,建立胶质瘤原位移植瘤模型,利用小鼠活体成像系统监测脑内肿瘤的生长情况,并通过石蜡切片,HE染色评价细胞在裸鼠脑内的病理特征及成瘤能力.结果 成功构建稳定表达GFP荧光和luciferase荧光的U251胶质瘤细胞系及动物模型,慢病毒整合并未改变细胞的增殖、迁移及侵袭能力;模型生长周期适中,成瘤率高,瘤体在颅内生长稳定,HE切片符合人胶质瘤特征.结论 双荧光标记的胶质瘤细胞相比于传统细胞更有利于胶质瘤动物模型的实验研究;U251-GFP-Luc胶质瘤细胞裸鼠模型,其肿瘤生长和病理特性与人胶质瘤相似,且可实时观察肿瘤生长,可作为胶质瘤实验研究的理想动物模型.
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