Rag1、Rag2 基因缺陷小鼠的构建及在异种肿瘤移植中的应用

doi:10.3969/j.issn.1674-5817.2016.04.004

实验动物与比较医学 ›› 2016, Vol. 36 ›› Issue (4): 263-269.DOI: 10.3969/j.issn.1674-5817.2016.04.004

Rag1、Rag2 基因缺陷小鼠的构建及在异种肿瘤移植中的应用

沈如凌^1,2, 石嘉豪¹, 盛哲津¹, 冯晓龙², 吴文婷², 费俭^1,2

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

收稿日期:2016-03-10 出版日期:2016-08-25 发布日期:2016-08-25
作者简介:沈如凌(1981-), 女, 助理研究员, 研究方向: 肿瘤学。E-mail: alieen_shen@163.com
基金资助:
上海市科委科研计划项目(14431904600, 13DZ2280600)

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

SHEN Ru-ling^1,2, SHI Jia-hao¹, SHENG Zhe-jin¹, FENG Xiao-long², WU Wen-ting², FEI Jian^1,2

1. School of Life Science and Technology, Tongji University, Shanghai 200092, China;
2. Shanghai Research Center for Model Organisms, Shanghai 201210, China

Received:2016-03-10 Online:2016-08-25 Published:2016-08-25

摘要/Abstract

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。

关键词: 重组激活基因(Rag1基因), Rag2基因, 基因缺陷, 免疫缺陷

Abstract: Objective To establish recombination-activating genes, Rag1, 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 Rag1 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 Rag1 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 Rag1, Rag2 mutant mice than that in BALB/c nude mice and NOD-SCID mice. Conclusion Rag1, Rag2 deficiency mouse models were successfully developed. Two mouse models display serious immune system defects, which could be used as exogenous tumor transplant recipients to establish tumor model.

Key words: Recombination-activating genes (Rag1 gene), Rag2 gene, Gene defect, Immunodeficiency

中图分类号:

Q95-33

沈如凌, 石嘉豪, 盛哲津, 冯晓龙, 吴文婷, 费俭. Rag1、Rag2 基因缺陷小鼠的构建及在异种肿瘤移植中的应用[J]. 实验动物与比较医学, 2016, 36(4): 263-269.

SHEN Ru-ling, SHI Jia-hao, SHENG Zhe-jin, FENG Xiao-long, WU Wen-ting, FEI Jian. Generation of Rag1 and Rag2 Deficient Mice and Their Application in Tumor Xenograft[J]. Laboratory Animal and Comparative Medicine, 2016, 36(4): 263-269.

参考文献

[1] Fugmann SD, Lee AI, Shockett PE, et al.The RAG proteins and V(D)J recombination: complexes, ends, and transposition[J]. Annu Rev Immunol, 2000,18(1):495-527.
[2] Fugmann SD.RAG1 and RAG2 in V(D)J recombination and transposition[J]. Immunol Res, 2001, 23(1):23-39.
[3] Oettinger MA.Activation of V(D)J recombination by RAG1 and RAG2[J]. Trends Genet, 1992, 8(12):413-416.
[4] Schatz DG, Oettinger MA, Baltimore D. The V(D)J recombination activating gene, RAG-1[J]. Cell, 1989, 59(6):1035-1048.
[5] Agrawal A, Schatz DG.RAG1 and RAG2 form a stable postcleavage synaptic complex with DNA containing signal ends in V(D)J recombination[J]. Cell, 1997, 89(1):43-53.
[6] Ru H, Chambers MG, Fu TM, et al.Molecular mechanism of V(D)J recombination from synaptic RAG1-RAG2 complex structures[J]. Cell, 2015, 163(5):1138-1152.
[7] Wilson A, Held W, MacDonald HR. Two waves of recombinase gene expression in developing thymocytes[J]. J Exp Med,1994, 179(5):1355-1360.
[8] Yu W, Nagaoka H, Jankovic M, et al.Continued RAG expression in late stages of B cell development and no apparent re-induction after immunization[J]. Nature,1999, 400(6745):682-687.
[9] Shaw AC, Swat W, Ferrini R, et al.Activated Ras signals developmental progression of recombinase-activating gene (RAG)-deficient pro-B lymphocytes[J]. J Exp Med, 1999,189(1):123-129.
[10] Corneo B, Moshous D, Gungor T, et al.Identical mutations in RAG1 or RAG2 genes leading to defective V(D)J recombinase activity can cause either T-B-severe combined immune deficiency or Omenn syndrome[J]. Blood, 2001, 97(9):2772-2776.
[11] Mombaerts P, Iacomini J, Johnson RS, et al.RAG-1-deficient mice have no mature B and T lymphocytes[J]. Cell, 1992, 68(5):869-877.
[12] Shinkai Y, Rathbun G, Lam KP, et al.RAG-2-deficient mice lack mature lymphocytes owing to inability to initiate V(D) J rearrangement[J]. Cell, 1992, 68(5):855-867.
[13] Bentzien F, Struman I, Martini JF, et al.Expression of the antiangiogenic factor 16K hPRL in human HCT116 colon cancer cells inhibits tumor growth in Rag1(-/-) mice[J]. Cancer Res, 2001, 61(19):7356-7362.
[14] Shultz LD, Goodwin N, Ishikawa F, et al.Human cancer growth and therapy in NOD/SCID/IL2Rynull(NSG) mice[J]. Cold Spring Harb Protoc, 2014, 7(1):694-708.
[15] Naugler WE, Tarlow BD, Fedorov LM, et al.Fibroblast growth factor signaling controls liver size in mice with humanized livers[J]. Gastroenterology, 2015, 149(3):728-740.
[16] Christa TR, Andre F, Christian F, et al.Composition of intestinal microbiota in immune-deficient mice kept in three different housing conditions[J]. PLoS One, 2014, 9(11): e113406.
[17] Seymour RL, Adams AP, Leal G, et al.A rodent model of Chikungunya virus infection in RAG1-/- mice, with features of persistence, for vaccine safety evaluation[J]. PLoS One, 2015,6(26):e0003800.

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

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