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目的：利用体外转染绿色荧光蛋白基因的胶质瘤细胞大鼠移植瘤模型，研究胶质瘤体内侵袭行为。方法：携有增强型绿色荧光蛋白（EnhancedGreenFluorescentProtein，EGFP）基因的pEGFP-N3质粒体外转染C6胶质瘤细胞，筛选稳定表达绿色荧光蛋白的细胞克隆，并作流式细胞仪和电镜检测，以立体定向法植入SD大鼠脑实质内建立大鼠移植瘤模型。4周后处死大鼠并作鼠脑连续石蜡切片，相邻切片分别作苏木素—伊红（HE）或免疫组化染色后荧光显微镜（紫外激发波长为488nm）检测。结果：EGFP于体内稳定表达，在荧光显微镜下可较容易区分肿瘤区与非肿瘤区界限，并能发现侵袭至远处的单个肿瘤细胞，其敏感性及特异性明显优于HE染色或免疫组化方法。结论：绿色荧光蛋白体外转染C6胶质瘤细胞并建立大鼠脑内移植瘤模型，是研究胶质瘤体内侵袭和转移的有效方法。
OBJECTIVE: To investigate the invasive behavior of glioma in vivo by using a rat model of glioma cells transfected with the green fluorescent protein gene in vitro. METHODS: C6 glioma cells were transfected with pEGFP-N3 plasmid carrying Enhanced Green Fluorescent Protein (EGFP) gene in vitro, and cell clones stably expressing green fluorescent protein were screened for flow cytometry and electron microscopy. Stereotactic implantation of SD rat brain parenchyma to establish a rat model of transplanted tumors. After 4 weeks, the rats were sacrificed and consecutively paraffinized. The adjacent sections were examined with hematoxylin-eosin (HE) or immunohistochemical staining (UV-excitation wavelength: 488 nm). RESULTS: EGFP was stably expressed in vivo. It was easier to distinguish the boundaries between tumor and non-tumor areas by fluorescence microscopy, and it was possible to find single tumor cells that invaded far away. Their sensitivity and specificity were significantly better than those of HE staining or immunization group. Method. Conclusion: Transfecting C6 glioma cells with green fluorescent protein in vitro and establishing a model of intracerebral transplantation in rats is an effective method to study invasion and metastasis of glioma in vivo.