荧光素酶标记小鼠结肠癌细胞建立活体成像移植瘤模型  

Establishment of a live imaging xenograft model by luciferase-labeled mouse colon cancer cells

作　　者：罗华灶 李雪 依含 谢君 朱乃硕 LUO Hua-Zao;LI Xue;YI Han;XIE Jun;ZHU Nai-Shuo(Laboratory of Microbial Infection and Molecular Immunology,School of Life Sciences,Fudan University,Shanghai 200438,China)

机构地区：[1]复旦大学生命科学学院微生物感染与分子免疫学实验室,上海200438 [2]复旦大学生物医学研究院,上海200032

出　　处：《中国免疫学杂志》2019年第17期2106-2111,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金面上项目(81472843);上海市科学技术委员会上海市科研计划项目(14ZR1424700)资助

摘　　要：目的:建立稳定表达荧光素酶的小鼠结肠癌细胞株并构建适用于小动物活体成像系统观察的BALB/c鼠皮下移植瘤模型。方法:通过基因重组的方式,将外源基因插入到逆转录病毒载体上,以工具细胞293T包毒后得到病毒,将携带荧光素酶基因的质粒转染到小鼠结肠癌细胞株CT26中,Puromycin筛选出稳定表达荧光素酶的单克隆细胞株。扩增后接种于BALB/c鼠,建立BALB/c鼠皮下移植瘤模型,通过活体动物成像系统监测肿瘤的生长过程。结果:获得了高水平稳定表达荧光素酶的结肠癌单克隆细胞株,该单克隆细胞株与母细胞系CT26具有相似的生长特性。将稳定表达荧光素酶的克隆接种于BALB/c鼠皮下可成瘤,小动物活体成像系统能准确监测肿瘤细胞在体内的生长过程。结论:成功建立了稳定表达荧光素酶的结肠癌单克隆细胞株。采用活体动物成像系统构建的BALB/c鼠皮下结肠癌移植瘤模型是拓展肿瘤体内生长、转移及治疗相关研究的理想模型。Objective: To obtained a mouse colon cancer cell strain stably expressing luciferase and a BALB/c mouse subcutaneous transplantation tumor model suitable for observation in a small animal living imaging system. Methods: The exogenous gene was inserted into the retroviral vector by gene recombination,and the virus was obtained by inoculating the 293T cells.The plasmid carrying the luciferase gene was transfected into mouse colon cancer cell line CT26,a monoclonal cell strain stably expressing luciferase was screened by using puromycin.After amplification,they were inoculated into BALB/c mice to establish a BALB/c mouse subcutaneous tumor model,and the tumor progression was monitored by a living animal imaging system. Results: A colon monoclonal cell strain stably high expressing luciferase was obtained,which had similar growth characteristics to wild-type CT26.The CT26-LUC1 stably expressing luciferase was inoculated into the subcutaneous of BALB/c mice to establish a tumor model,and the small animal living imaging system can accurately monitor the progression of tumor cells in vivo. Conclusion: A colon monoclonal cell strain stably expressing luciferase was successfully established.The construction of a BALB/c mouse subcutaneous colon xenograft model suitable for living animal imaging systems is an ideal model for expanding the scope of research on progression,metastasis and treatment of tumors in vivo.

关 键 词：荧光素酶 CT26细胞 移植瘤 活体成像 BALB/C鼠 

分 类 号：R739.6[医药卫生—肿瘤]