Rag1、Rag2基因缺陷小鼠的构建及在异种肿瘤移植中的应用  

Generation of Rag1 and Rag2 Deficient Mice and Their Application in Tumor Xenograft

作　　者：沈如凌[1,2] 石嘉豪[1] 盛哲津[1] 冯晓龙 吴文婷 费俭[1,2] SHEN Ru-ling SHI Jia-hao SHENG Zhe-jin FENG Xiao-long WU Wen-ting FEI Jian(School of Life Science and Technology, Tongji University, Shanghai 200092, China Shanghai Research Center for Model Organisms, Shanghai 201210, China)

机构地区：[1]同济大学生命科学与技术学院,上海200092 [2]上海南方模式生物研究中心,上海201210

出　　处：《实验动物与比较医学》2016年第4期263-269,279,共8页Laboratory Animal and Comparative Medicine

基　　金：上海市科委科研计划项目（14431904600,13DZ2280600）

摘　　要：目的 建立重组激活基因,Rag1和Rag2基因缺陷小鼠模型,并对其表型和异种肿瘤移植的成瘤性进行分析.方法 通过传统的胚胎干细胞（ES）细胞打靶、囊胚注射方式,分别构建Rag1和Rag2基因缺陷小鼠模型;通过流式细胞术对缺陷小鼠外周血T/B淋巴细胞含量进行检测;通过外源异种肿瘤细胞移植实验,对移植肿瘤细胞的成瘤性进行评价.结果 建立了Rag1和Rag2基因缺陷小鼠模型,两种基因突变纯合子小鼠模型外周血中T、B淋巴细胞含量较野生型小鼠显著降低;人源肿瘤细胞A549在上述两种基因突变纯合子小鼠中较BALB/c-nu（裸小鼠）或（NOD-SCID）非肥胖糖尿病/重症联合免疫缺陷小鼠表现出更强的成瘤性.结论 分别成功建立了Rag1和Rag2基因缺陷小鼠模型,两种小鼠模型均发生了T、B淋巴细胞生成障碍,导致免疫系统严重缺陷,可作为异种外源肿瘤移植的受体,用于小鼠荷瘤模型的建立.Objective To establish recombination-activating genes, Ragl, Rag2 deficiency mouse models and study the possibility as tumor xenograft recipients. Methods The traditional embryonic stem （ES） cell targeting and blastocyst microinjection were performed to develop the Ragl and Rag2 deficiency mouse models. The flow cytometry assay was conducted to analyze the T/B lymphocyte contents in peripheral blood. The tumorigenicity on different immunodeficiency mouse strains was assessed by exogenous tumor cell transplantation experiment. Results Ragl and Rag2 deficiency mouse models were established. The T/B lymphocytes of peripheral blood in two strains of homozygous mutant mouse models were significantly reduced compared with those of wild type mice. A549 tumor cells had stronger tumorigenicity in Ragl, Rag2 mutant mice than that in BALB/c nude mice and NOD- SCID mice. Conclusion Ragl, Rag2 deficiency mouse models were successfully developed. Two mouse models display serious immune system defects, which could be used as exogenous tumor trans- plant recipients to establish tumor model.

关 键 词：重组激活基因(Rag1基因) Rag2基因 基因缺陷 免疫缺陷 Recombination-activating GENES (Ragl gene) 

分 类 号：R73-3[医药卫生—肿瘤]