绿色荧光蛋白标记的大鼠C6胶质瘤模型的构建

Establishment of rat C6 glioma model engineered by green fluorescent protein

目的探讨绿色荧光蛋白(GFP)标记的大鼠C6胶质瘤模型的构建方法并观察其作用。方法用带有增强型GFP(EGFP)基因的p EGFP-C1质粒转染C6细胞,通过G418筛选获得稳定表达EGFP的C6细胞克隆EGFPC1-C6细胞。采用立体定向法,将浓度为1×106/L的EGFP-C1-C6细胞悬液10μL注入16只SD大鼠的颅内,建立EGFP标记的大鼠C6胶质瘤模型。观察大鼠接种肿瘤细胞后的反应及存活时间。分别于造模后第7、14、21天对大鼠行头颅MRI检查,计算肿瘤体积。常规HE染色观察病理学形态,激光共聚焦显微镜下观察胶质瘤的浸润及侵袭情况。结果成功培养了具有EGFP标记的EGFP-C1-C6细胞,MRI及病理学检查发现所有大鼠颅内均有肿瘤生长,肿瘤体积为(244.60±36.06)mm3,采用激光共聚焦显微镜观察易于发现经EGFP标记的肿瘤组织,肿瘤区域清晰可见,并较易发现单个肿瘤细胞的远处浸润生长。结论采用EGFP-C1-C6细胞成功构建了大鼠C6胶质瘤模型,该模型有助于显示胶质瘤细胞的侵袭、转移及微小侵袭灶。

Objective To establish the rat C6 glioma model engineered by green fluorescent protein and explore its effect. Methods C6 cells were transfected with p EGFP-C1 plasmid expressing enhanced green fluorescent protein（ EGFP） gene. Then the transfected cells were selected by G418 to obtain the stable C6 cell clones（ p EGFP-C1-C6）. p EGFPC1-C6 cell suspension was injected into intracal of 16 SD rats to establish rat C6 glioma model with green fluorescent protein by stereotaxic apparatus. After inoculation,cell life cycle was observed and tumor growth status was dynamically documented by magnetic resonance imaging. At 7,14,21 day after glioma model established,glioma tumor volumes were detected and measured by MRI. Brain specimens were observed by HE staining. Confocal laser scanning microscopy was used to observe the infiltration and invasion of glioma cell. Results Rat C6 glioma model with green fluorescent protein was established successfully. Tumor cells invasion can be easily detected by green fluorescent protein imaging and MRI. Tumor volume was（ 244. 60 ± 36. 06） mm^3. It was easier to observe the micro-metastasis and invasive. Conclusions Rat C6 glioma model could be successful established with EGFP-C1-C6 cells,which was more sensitive in showing invade,metastasis and single invasive tumor cell.