人急性早幼粒细胞白血病小鼠模型的建立

中国临床药理学与治疗学 ›› 2013, Vol. 18 ›› Issue (8): 858-863.

人急性早幼粒细胞白血病小鼠模型的建立

王骏^1,2, 王珊珊², 杨德宣², 袁守军²

¹安徽医科大学研究生院,合肥 230032,安徽;
²军事医学科学院放射与辐射医学研究所药理毒理研究室,北京 100850

收稿日期:2013-01-17 修回日期:2013-07-12 出版日期:2013-08-26 发布日期:2013-07-31
通讯作者: 袁守军,男,教授,硕士/博士生导师,研究方向:肿瘤药理学。Tel: 010-66930271 E-mail: yuansj@nic.bmi.ac.cn
作者简介:王骏,男,硕士研究生,研究方向:肿瘤药理学。Tel: 010-66931215 E-mail: wangj.521@163.com
基金资助:
重大新药创制国家科技重大专项综合性新药研究开发技术大平台项目(2012ZX09301003-001)

Development of a murine model of acute promyelocytic leukemia

WANG Jun^1,2, WANG Shan-shan², YANG De-xuan², YUAN Shou-jun²

¹The Graduate School of Anhui Medical University, Hefei 230032, Anhui,China;
²Department of Pharmacology, Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China

Received:2013-01-17 Revised:2013-07-12 Online:2013-08-26 Published:2013-07-31

摘要/Abstract

摘要： 目的: 探索稳定、可靠且重复性较好的人急性早幼粒细胞白血病Nod/SCID小鼠模型的建立方法,并观察模型的病程特点。方法: Nod/SCID小鼠经⁶⁰Co γ射线照射,照后第5天,外周血白细胞降至较低水平,此时尾静脉注射接种HL60细胞。通过流式细胞计数分析外周血中HL60细胞比例,以观察白血病细胞在外周血中分布变化;日常观察小鼠身体浅表部位实体瘤的形成及生长;并应用组织病理学方法确认组织脏器中肿瘤播散状况。结果: 接种第7天,Nod/SCID小鼠外周血中可检测到HL60细胞;接种 30 d 左右,可观察到小鼠出现全身播散的实体瘤;40 d 左右,动物体质状况显著衰退,小鼠陆续死亡。小鼠的造血系统及非造血系统的组织及脏器均出现肿瘤的浸润。重复实验结果一致。结论: Nod/SCID小鼠经⁶⁰Co γ射线照射后第5天,尾静脉注射接种HL60细胞,小鼠的病程可控制在 40 d 左右,且该模型符合临床疾病发展规律,可应用于骨髓毒性较低的受试药物的药效筛选及评价。

关键词: 小鼠模型, 早幼粒细胞白血病, HL60细胞, Nod/SCID小鼠

Abstract: AIM: To explore a method to develop a stable and repeatable murine model of acute promyelocytic leukemia(APL),which progression could be observed.METHODS: For the model, Nod/SCID mice were irradiated by ⁶⁰Co γ ray and the level of leukocytes was very low at the fifth day, then Nod/SCID mice were injected with HL60 cells intravenously via tail vein.The percentage of HL60 cells in the peripheral blood was determined by flow cytometry, which was used to observe the distribution change of HL60 cells in the peripheral blood.The development of solid tumors in the superficial parts of body was observed daily, and to confirm the characteristics of tumor spread in the mice with histopathological methods.RESULTS: Day 7 post injection, HL60 cells could be detected in the peripheral blood of Nod/SCID mice. About day 30 post injection, solid tumors were observed through the whole body of mice. About day 40, mouse physical fitness began to decline, and mice began to die.The leukemic infiltration was found in the hematopoietic system and non-hematopoietic system. The results of two trials were coincidence.CONCLUSION: For the model, Nod/SCID mice were irradiated by γ ray from ⁶⁰Co, injected with HL60 cells intravenously via tail vein in the day 5 post irradiation. The course of disease could be kept in about 40 days, and leukemic progression in the Nod/SCID mice with APL was similar with clinical leukemic progression.The model could be used for anti-leukemia drug screening and pharmacodynamic evaluation of candidate drugs with low bone marrow toxicity.

Key words: Murine model, Acute promyelocytic leukemia, HL60 cell , Nod/SCID mice

中图分类号:

R965.2

王骏, 王珊珊, 杨德宣, 袁守军. 人急性早幼粒细胞白血病小鼠模型的建立[J]. 中国临床药理学与治疗学, 2013, 18(8): 858-863.

WANG Jun, WANG Shan-shan, YANG De-xuan, YUAN Shou-jun. Development of a murine model of acute promyelocytic leukemia[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2013, 18(8): 858-863.

参考文献

[1] 周永列,吕亚萍,胡惟孝,等.四嗪二甲酰胺对白血病细胞株HL-60 体外作用的研究[J].中国临床药理学与治疗学,2006,11(9):1006-1012.
[2] 阮晶晶,陈飞虎,徐佼,等.新型维甲酸衍生物诱导NB4 细胞分化的初步研究[J].中国临床药理学与治疗学,2008,13(11):1237-1242.
[3] 吴建国,周永列,夏大静,等.大萼香茶菜甲素通过线粒体途径诱导HL-60 白血病细胞凋亡[J].中国临床药理学与治疗学,2009,14(7):741-747.
[4] Roomi MW, Roomi NW, Bhanap B,et al. Nutrient mixture inhibits in vitro and in vivo growth of human acute promyelocytic leukemia HL-60 cells [J]. Exp Oncol, 2011, 33(4): 212-215.
[5] Liu W, Zhu YS, Guo M, et al. Therapeutic efficacy of NSC606985, a novel camptothecin analog, in a mouse model of acute promyelocytic leukemia [J]. Leuk Res, 2007, 31(11): 1565-1574.
[6] Honma Y, Ishii Y, Sassa T, et al. Treatment of human promyelocytic leukemia in the SCID mouse model with cotylenin A, an inducer of myelomonocytic differentiation of leukemia cells [J]. Leuk Res, 2003, 27(11): 1019-1025.
[7] Lewis ID, McDiarmid LA, Samels LM, et al. Establishment of a Reproducible Model of Chronic-Phase Chronic Myeloid Leukemia in Nod/SCID Mice Using Blood-Derived Mononuclear or CD₃₄⁺Cells [J]. Blood, 1998, 91(2): 630-640.
[8] Tomkinson B, Bendele R, Giles FJ, et al. OSI-211, a novel liposomal topoisomerase I inhibitor, is active in SCID mouse models of human AML and ALL [J]. Leuk Res, 2003, 27(11): 1039-1050.
[9] Chun-Guang W, Jun-Qing Y, Bei-Zhong L, et al. Anti-tumor activity of emodin against human chronic myelocytic leukemia K562 cell lines in vitro and in vivo[J]. Eur J Pharmacol,2010, 627(1/2/3): 33-41.
[10] Liem NLM, Papa RA, Milross CG, et al. Characterization of childhood acute lymphoblastic leukemia xenograft models for the preclinical evaluation of new therapies [J]. Blood, 2004, 103(10):3905-3914.
[11] Benito J, Shi Y, Szymanska B,et al. Pronounced Hypoxia in Models of Murine and Human Leukemia: High Efficacy of Hypoxia-Activated Prodrug PR-104 [J]. PLoS ONE, 2011, 6(8): e23108.
[12] Liu Y,Li LJ,Tang,et al. Treatment of lycorine on SCID mice model with human APL cells[J]. Biomed Pharmacother, 2007, 61(4): 229-234.
[13] Ram'rez M, D'az MA, Madero L, et al. Transplantation of marrow cells from children with standard risk-acute lymphoblastic leukemia at the end of therapy into Nod/SCID mice for detecting residual leukemic cells with in vivo growth potential [J]. Leuk Res, 2003, 27(12): 1153-1157.
[14] Shankar DB, Li JL, Tapang P, et al. ABT-869, a multitargeted receptor tyrosine kinase inhibitor: inhibition of FLT3 phosphorylation and signaling in acute myeloid leukemia [J]. Blood, 2007, 109(8): 3400-3408.
[15] Chao MP, Alizadeh AA, Tang C, et al. Therapeutic antibody targeting of CD47 eliminates human acute lymphoblastic leukemia [J]. Cancer Res, 2011, 71(4): 1374-1384.
[16] Zhang Z, Zhang ML, Garmestani K, et al. Effective treatment of a murine model of adult T-cell leukemia using 211 At-7G7/B6 and its combination with unmodified anti-Tac (daclizumab) directed toward CD25 [J]. Blood, 2006, 108(3): 1007-1012.
[17] Nijmeijera BA, Mollevanger P, Zelderen-Bhola SL, et al. Monitoring of engraftment and progression of acute lymphoblastic leukemia in individual Nod/SCID mice [J]. Exp Hematol,2001, 29(3): 322-329.