GFP和Luc双标技术在小鼠肿瘤模型建立中的应用  

Application of GFP/Luc Double-tagging Technique in Tumour Model of Mouse

作　　者：王晓敏[1] 闫明霞[1] 梁琳慧[1] 李金枝[2] 朱淼鑫[1] 吴一迁[1] 姚明[1] 

机构地区：[1]上海交通大学肿瘤研究所,上海200032 [2]复旦大学附属金山医院,上海200540

出　　处：《实验动物与比较医学》2010年第1期2-7,共6页Laboratory Animal and Comparative Medicine

基　　金：上海市科委创新行动计划（08140902400和071409009）

摘　　要：目的建立GFP（绿色荧光蛋白）／Luc（荧光素酶）双标高效表达的人肺癌细胞系和人肝癌细胞系，利用活体成像系统观察裸小鼠肺原位移植瘤和肝原位移植瘤的生长情况。方法应用慢病毒转染的方法建立GFP／Luc双标的SPC-A-1和SMMC-7721细胞系，结合流式细胞分选技术使GFP／Luc双标表达率接近100％，分别接种于裸小鼠肺原位和肝原位，采用活体成像技术监测各稳定转染细胞在小鼠体内深部组织的表达情况。结果成功建立了GFP／Luc双标表达率接近100％的SPC-A-1-GFP／LHC和SMMC-7721-GFP／Ltic细胞系，进行裸小鼠肺原位移植和肝原位移植后，应用活体成像系统观测到裸小鼠肺原位和肝原位早期的微小病灶和晚期移植瘤的生长情况，而且，随着肿瘤移植时间的延长，移植瘤体积的增大，活体成像检测到的发光面积逐渐增大，发光强度也逐渐增加。结论所建立的稳定表达GFP／Luc的细胞系，结合活体成像技术，能够对动物模型深部的肿瘤病灶进行活体、实时、定量检测，且灵敏度极高，为进一步研究肿瘤的发生及发展提供了理想的技术手段。Objective To establish human lung cancer cell line and human liver cancer cell line with GFP/Luc double-tagging which having high expressing rate, and to observe development of the tumors in deep organisms of the nude mice with in vivo biofluorescence imaging. Methods The SPC-A-1- GFP/Luc cell line and SMMC-7721-GFP/Luc cell line were established by lentivirus transfection, enhancing their GFP/Luc double-tagging expressing rate to about 100% by flow cytometer, then were inoculated in lung and liver of the BALB/c nude mice respectively, finally, the behaviors of the two cell lines transfected stably in development of the tumours was detected by in vivo biofluorescence imaging. Results The cell lines of SPC-A-1-GFP/Luc and SMMC-7721-GFP/Luc with GFP/Luc double-tagging have been built up with the expressing rate nearly 100%. After the two cell lines have been inoculated in lung and liver of the nude mice, the tiny tumor focuses in early time and the status of tumors in late time in the deep organisms were observed by the in vivo biofluorescence imaging. Conclusion The established cell lines with stable GFP/Luc double-tagging, coupled with the application of in vivo biofluorescence imaging, can detect the tumors in deep organisms of the nude mouse intravitaly, real-timely, quantitively, and may be offering an ideal device for researchers to further explore development of the tumor.

关 键 词：GFP/Luc双标 慢病毒载体 活体成像系统 裸小鼠 肿瘤模型 

分 类 号：R73-3[医药卫生—肿瘤]