SHG-44绿色荧光裸鼠原位模型的建立  

Establishment of orthotopic model of SHG-44 in enhanced green fluorescent protein nude mice

作　　者：陈雄辉[1] 周幽心[1] 谢学顺[1] 陈金明[1] 苏作鹏[1] 

机构地区：[1]苏州大学附属第一医院神经外科暨脑神经研究室,215006

出　　处：《中华实验外科杂志》2012年第5期966-968,F0004,共4页Chinese Journal of Experimental Surgery

基　　金：江苏省卫生厅135重点学科建设基金资助项目（XK200-722）;苏州市科技计划资助项目（SYS201025）

摘　　要：目的建立SHG-44绿色荧光裸鼠原位模型。方法将C57BL／6J-增强绿色荧光蛋白（EGFP）转基因小鼠与裸小鼠进行交配，在继代时严格挑选都表达EGFP基因的无毛雄鼠（6）与有毛母鼠（早）交配，通过肉眼、荧光显微镜和分子生物学等方法观察EGFP基因在裸小鼠全身各组织中的表达；将胶质瘤细胞株SHG-44（15μl，含1×10^5个细胞）注射于绿色荧光裸鼠的脑内制成原位模型。结果每只种鼠平均产仔7～8只，其中纯合裸仔鼠有3～4只，比例接近1：1；每代仔鼠通过小动物活体成像系统出生后2d就可鉴别其是否表达EGFP基因；器官及组织冰冻切片荧光显微镜下观察均见有绿色荧光；聚合酶链反应（PCR）结果表明脑组织、皮下肌肉、鼠尾和脚趾全部表达EGFP基因；原位模型的成瘤率达100％，瘤细胞依然具有人脑胶质瘤的基本特征。结论表达EGFP基因的裸小鼠可用于肿瘤移植实验。Objective To establish the orthotopic model of SHG-44 in the enhanced green fluores- cent protein （EGFP） nude mice. Methods The C57BL/6J-EGFP transgenic mice were crossed with nude mice, then the hairless male nude mouse expressing EGFP was crossed with the hairy female pubescence mouse to obtain the EGFP nude mouse. The expression status of EGFP in the EGFP nude mice was evaluated under the fluorescence microscopy and by using polymerase chain reaction （PCR）. The human glioma cell line SHG-44 cells （ 15 μL cell suspensions, including 1 ×10^6 cells） were stereotaxically implanted into the brain of the EGFP nude mouse to produce orthotopic models. Results Each mouse could give birth to 7-8 litter size on average, and there were 3-4 naked young homozygous mice, the ratio close to 1： 1. Through the small animal in vivo imaging system 2 days after birth, the expression of EGFP gene could be identified. Green fluorescence was seen in the frozen sections of organs and tissues under the fluorescent microscopy. PCR results showed that the brain tissue, subcutaneous muscle, tail and toes all expressed the EGFP gene. The tumor formation rate of the orthotopic model was about 100%. The cells of orthotopic model still had the basic characteristics of human glioma cells. Conclusion The EGFP nude mouse can be used for human tumors transplantation experiments.

关 键 词：绿色荧光蛋白 裸鼠 转基因动物 

分 类 号：R73-35[医药卫生—肿瘤] R-332[医药卫生—临床医学]