应用NB4细胞株建立SCID beige小鼠急性早幼粒细胞白血病病理模型

发布时间：2019-05-21 14:25

【摘要】：目的:本研究旨在建立稳定、有效、重复性好的人急性早幼粒白血病重度联合免疫缺陷(SCID)小鼠移植瘤模型,观察模型的病程特点和生物学行为。方法:将3-5周龄SCID beige小鼠随机分为实验组和对照组;实验组鼠尾静脉注射对数生长期的NB4细胞5×106个/只,动态监测小鼠外周血白细胞数和外周血涂片中人早幼粒细胞阳性率;应用形态学和组织病理学方法确认白血病细胞浸润肝、脾、肺、肾、脑等器官情况,Western blot检测组织中PML-RARa融合蛋白表达情况。结果:接种NB4细胞2周内的实验组SCID小鼠外周血白细胞计数与对照组相比差异无统计学意义(P0.05);同时,血涂片姬姆萨染色后镜检未见到NB4细胞;接种第21天和第28天实验组SCID小鼠外周血白细胞计数分别为(4.79±1.13)×109/L和(7.62±2.24)×109/L,显著高于对照组(P0.05);血涂片吉姆萨染色后可见NB4细胞比例分别占(2.14±0.63)%和(6.6±2.76)%;形态学观察显示,接种21 d后实验组SCID小鼠体内出现单个或多个肿瘤实体包块;同时,组织切片HE染色表明,肝、脾、肺、肾、脑组织均有不同程度的瘤细胞浸润;细胞免疫荧光结果显示,实验组小鼠骨髓细胞CD33表达呈阳性(P0.05);Western blot数据证实,肝、肾、脑组织中检测到不同水平的PML-RARa融合蛋白表达。结论:SCID小鼠鼠尾静脉注射NB4细胞可成功构建人急性早幼粒细胞白血病小鼠模型,该模型能模拟临床白血病累及骨髓和弥漫生长特点,是研究人类白血病发病机理及实验治疗的良好模型。
[Abstract]:Objective: to establish a stable, effective and reproducible human acute promyelocytic leukemia severe combined immunodeficient (SCID) mouse model, and to observe the course of disease and biological behavior of the model. Methods: SCID beige mice aged 3 to 5 weeks were randomly divided into experimental group and control group. In the experimental group, 5 脳 10 ~ 6 NB4 cells were injected intravenously into the tail vein to dynamically monitor the number of peripheral blood leukocytes and the positive rate of human progranulocytes in peripheral blood smears. The expression of PML-RARa fusion protein in leukemic cells infiltrated into liver, spleen, lung, kidney and brain was detected by morphology and histology., Western blot was used to detect the expression of PML-RARa fusion protein in liver, spleen, lung, kidney, brain and other organs. Results: there was no significant difference in peripheral blood leukocyte count between the experimental group and the control group within 2 weeks after inoculated with NB4 cells (P 0.05). At the same time, no NB4 cells were found in the blood smear after Giemsa staining. On the 21st and 28th day of vaccination, the peripheral blood leukocyte counts of SCID mice in the experimental group were (4.79 卤1.13) 脳 109 脳 L and (7.62 卤2.24) 脳 109 / L, respectively, which were significantly higher than those in the control group (P 0.05). The percentage of NB4 cells in blood smears was (2.14 卤0.63)% and (6.6 卤2.76)%, respectively, and the morphological observation showed that one or more tumor solid masses appeared in SCID mice in the experimental group 21 days after vaccination, and the percentage of DNA cells in the experimental group was (2.14 卤0.63)% and (6.6 卤2.76)%, respectively. At the same time, HE staining showed that there were different degrees of tumor cell infiltration in liver, spleen, lung, kidney and brain tissue, and the results of cellular immunofluorescence showed that the expression of CD33 in bone marrow cells of the experimental group was positive (P 0.05). Western blot data confirmed that different levels of PML-RARa fusion protein expression were detected in liver, kidney and brain tissue. Conclusion: the mouse model of human acute promyelocytic leukemia can be successfully established by injecting NB4 cells into the tail vein of SCID mice. The model can simulate the characteristics of clinical leukemia involving bone marrow and diffused growth. It is a good model to study the pathogenesis and experimental treatment of human leukemia.
【作者单位】：西安交通大学医学院第一附属医院血液内科;
【基金】：陕西省科技统筹创新工程计划项目(2012KTCL03-12)
【分类号】：R733.7;R-332

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