尾静脉注射HL60、HL60/ADR细胞建立SCID beige小鼠白血病动物模型的研究  

Establishment of a human acute leukemia pathological model in SCID beige mice using HL60 and HL60/ADR cell lines

作　　者：张坤[1] 徐昊淼 程涓[1] 赵勇[2] 许亚梅[1] ZHANG Kun;XU Haomiao;CHENG Juan;ZHAO Yong;XU Yamei(Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China;Institute of Zoology,Chinese Academy of Sciences,Beijing 100101)

机构地区：[1]北京中医药大学东直门医院,北京100700 [2]中国科学院动物所,北京100101

出　　处：《中国比较医学杂志》2018年第7期43-51,共9页Chinese Journal of Comparative Medicine

基　　金：北京市自然科学基金(编号:7152092)

摘　　要：目的应用急性早幼粒细胞HL60及耐阿霉素(adriamycin,ADR)细胞HL60/ADR,构建SCID beige小鼠白血病模型。方法将15只4~5周龄的雌性SCID beige小鼠,随机分为对照组3只、四组模型12只(每组3只)。经2Gy X射线预处理,模型组分别经尾静脉接种对数生长期HL60细胞悬液1×10~7个/只(M1组)、5×10~6个/只(M2组),HL60/ADR细胞悬液1×10~7个/只(M3组)、5×10~6个/只(M4组),对照组鼠尾静脉注射等剂量生理盐水。观察一般情况,于照射前、造模第10、18、28天检测血常规、外周血白细胞分类、外周血CD33阳性细胞比例,第32天或濒死时处死取材,组织切片病理检查。结果各模型组于接种细胞第7天开始,出现竖毛、萎靡少动、脊背弓起、步态不稳、偏侧或转圈,M1、M3两组较M2、M4两组体重下降更明显(P<0.05),显著低于对照组(P<0.01)。至观察周期32 d,M1、M3两组生存率分别为33%、67%,M2、M4两组全部生存。各组小鼠经X线照射7 d内白细胞数迅速降低,随时间推移逐渐上升,第28天各模型组白细胞计数均显著高于对照组(P<0.01);且各模型组白血病细胞比例亦明显升高,以接种HL60/ADR的M3组最为显著(P<0.05);接种第28天时各模型组CD33阳性细胞比例显著高于对照组(P<0.01),然各模型组间差异无显著性。第32天或濒死前取材,病理组织切片,造模各组脾均可见弥漫性白血病细胞浸润,肝偶见白血病细胞浸润灶。结论 SCID beige小鼠经2Gy X射线预处理后,每只尾静脉注射HL60、HL60/ADR细胞1×10~7个或5×10~6个均可构建急性髓系白血病动物模型,符合急性髓系白血病的生物学特点,高浓度成瘤更快,中位生存期约29 d。Objective To establish a stable,effective,and reproducible acute promyelocytic leukemia model in severe combined immunodeficient(SCID)beige mice using acute myelocyte HL60 and adriamycin(ADR)-resistant HL60/ADR cell lines.Methods Female SCID beige mice(4–5 weeks old)were divided randomly into one control group and four model groups with three mice in each group.After X-ray irradiation,SCID mice received 1×10 7(M1 group)or 5×10 6(M2 group)HL60 cells,or 1×10 7(M3 group)or 5×10 6(M4 group)HL60/ADR cells via tail vein injection.The white blood cell(WBC)count and positive rate of promyelocytes in peripheral blood were dynamically monitored by detecting cells expressing CD33 using flow cytometry.Morphological examination and histopathological assays were employed to confirm promyelocyte infiltration into organs(liver,spleen,and kidneys).Results The four model groups displayed abnormal behaviors of tremors,retardation,and piloerection.The two high dose model groups experienced significant weight loss compared with the two low dose model groups(P<0.05).Moreover,the four model groups had significantly lower body weights than the control group(P<0.01).At day 32,survival rates of M1 and M3 groups were 33%and 67%,respectively.All mice in M2 and M4 groups survived.The WBC count in peripheral blood declined after X-ray irradiation.At day 28 after inoculation,the peripheral blood WBC counts of the four model groups were significantly higher than that in the control group(P<0.01).At day 28,the leukemic cell percentage of the M3 group was the highest compared with the other three model groups(P<0.05).The rates of CD33-positive cells in flow cytometric analysis of the four model groups were higher than that in the control group at day 28,but there were no significant differences between the four model groups.Morphological examination and HE staining of tissue biopsies demonstrated a large number of promyelocytes in the spleen and liver.Conclusions The human acute leukemia SCID beige mouse model was successfully established

关 键 词：小鼠模型 白血病 HL60细胞 HL60/ADR细胞 

分 类 号：R-33[医药卫生]