CB6F1小鼠急性粒单核细胞白血病M4(AML-M4)模型的建立及鉴定  

Reproduction and appraisal of an animal model of acute myelomonocytic leukemia in the CB6F1generation mice

作　　者：左洪莉[1] 彭恩兰[1] 刘铁强[1] 黄珊[1] 李玉芳[1] 姚晓兰[2] 尚世臣[2] 艾辉胜[1] 

机构地区：[1]解放军307医院血液科,北京100071 [2]军事医学科学院实验动物中心,北京100071

出　　处：《解放军医学杂志》2011年第1期53-57,共5页Medical Journal of Chinese People's Liberation Army

摘　　要：目的建立CB6F1小鼠急性粒单核细胞白血病M4(AML-M4)模型并进行鉴定。方法取CB6F1小鼠,经静脉注射小鼠粒单核细胞白血病WEHI-3细胞,观察不同数量(1×106、2×106、5×106、1×107个)细胞接种与小鼠生存时间的相关性。取接种1×106个细胞的小鼠为研究对象,动态观察其发病情况并在接种后每周行血常规检测。选择接种后4周因白血病发病濒死小鼠,取外周血行细胞涂片形态学观察、免疫表型检测及白血病细胞主要组织相容性复合体(MHC)检测,分离骨髓组织行细胞形态学、免疫化学和染色体核型分析,同时取肝、脾、肾、肺、心脏和脑组织行病理学观察,综合上述检测指标判断白血病模型建立的有效性。经腹腔分别注射阿糖胞苷(Ara-C)50、100mg/kg,观察小鼠发病情况及生存期,判断该模型对化疗药物的敏感性。所有实验均取正常小鼠作对照。结果接种不同数量WEHI-3细胞的小鼠均在17～33d发生白血病而死亡,接种细胞数量与生存期呈负相关(r=-0.936,P<0.01)。接种1×106个细胞的小鼠生存期为25～33(28.50±1.87)d,接种后4周因白血病发病濒死的小鼠表现如下:外周血白细胞数目显著增加,最高可达81×109/L,与正常小鼠比较差异显著(P<0.05)。外周血涂片可见胞体较大、形态不规则的白血病细胞。骨髓细胞涂片可见大量原始和幼稚细胞,过氧化酶(POX)、特异性酯酶(SE)、非特异性酯酶(NSE)及氟化钠(NaF)抑制实验均阳性,符合AML-M4的特征;各器官病理切片中均可见大量白血病细胞弥漫性浸润;骨髓细胞内染色体为超二倍体;外周血CD4+/CD8+比值下降,表达C-KIT和MAC-3的细胞均较正常小鼠增多(P<0.05);MHC表型为H2Kb-H2Kd+的细胞比例升高,濒死时表型以H2Kb-H2Kd+为主。Ara-C可延长该白血病模型型小鼠的生存期,且100mg/kg剂量效优低于50mg/kg(P<0.05)。结论静脉接种WEHI-3白血病细胞可在CB6F1小鼠中成功构建AML-M4白血病模�Objective To reproduce an acute myelomonocytic leukemia M4（AML-M4）animal model with the CB6F1generation mice.Methods The CB6F1（BALB/c C57BL/6）mice were inoculated intravenously with different amounts（1×10^6,2×10^6,5×10^6,1 ×10^7）of WEHI-3cells,a cell line of myelomonocytic leukemia.The correlation between the animal survival and the inoculated amount was analyzed.The mice,inoculated with 1×10^6 cells,were selected for observation of leukemia onset,and sampled for routine blood test.Four weeks after inoculation,the peripheral blood was collected from moribund mice,morphological observation was made in blood smears,and immunophenotype and major histocompatibility complex（MHC）was determined;the marrow cells were collected for morphological observation,and immunochemical and karyotype analysas were made.The liver,spleen,kidney,lung,heart and brain were obtained for pathological observation.The results of all the observations and determinations were then comprehensively analyzed to evaluate the authenticity of the established AML-M4mice model.Ara-C,in a dosage of 50mg/kg or 100mg/kg,was intraperitoneally injected to the model mice for observation of the disease course and survival of the animals,and to evaluate the sensitivity of model mice to the chemotherapeutics.Normal mice were selected to serve as control in all the experiments.Results Mice inoculated with different amount of WEHI-3cells died of leukemia 17to 33days after inoculation,and a negative correlation between the inoculated amount and the survival time of animals was observed（r=-0.936,P〈0.01）.Those inoculated with 1×106 cells survived for 25～33（28.50±1.87）days.Four weeks after inoculation,the WBC counts of peripheral blood increased obviously with a peak value of 81×109/L,in the moribund leukemic mice,which was significantly different from that of normal control（P〈0.05）.Leukemia cells with larger size and irregular shape were observed in the blood smear.A large number of primary and immature cells were found in t

关 键 词：白血病 粒-单核细胞 急性 WEHI-3细胞 模型 动物 

分 类 号：R733.713[医药卫生—肿瘤]