白血病多药耐药小鼠模型的构建及鉴定  

Construction and identification of multidrug resistance mouse model with leukemia

作　　者：闫理想 姜静 杨向东[1] 李德冠[3] 史哲新[1] YAN Lixiang;JIANG Jing;YANG Xiangdong(Department of Hematology,First Affiliated Hospital,Tianjin University of Traditional Chinese Medicine,Tianjin 300381,CHINA)

机构地区：[1]天津中医药大学第一附属医院血液科(国家中医针灸临床医学研究中心),300381 [2]天津市中医药研究院附属医院消化科 [3]中国医学科学院放射医学研究所

出　　处：《江苏医药》2023年第1期1-4,F0002,共5页Jiangsu Medical Journal

基　　金：天津市科技计划项目(21JCQNJC01210);天津中医药大学第一附属医院“拓新工程”基金项目(院2020057)。

摘　　要：目的探讨白血病多药耐药(MDR)小鼠模型的构建方法并对其进行鉴定。方法常规培养白血病MDR细胞系K562/A02,采用免疫磁珠法分选CD34^(+)CD38^(-)细胞亚群,流式细胞术检测CD34^(+)CD38^(-)细胞比例,CCK^(-)8法检测CD34^(+)CD38^(-)细胞亚群耐药性。采用移植瘤皮下包埋方法构建白血病MDR小鼠模型(A组,10只),同时设注射生理盐水的正常对照组(C组,10只)。记录小鼠肿瘤体积生长曲线及外周血细胞计数,流式细胞术检测小鼠骨髓细胞中CD34^(+)CD117^(+)细胞比例,CCK^(-)8法检测移植瘤组织细胞耐药性,HE染色观察肝脏、脾脏及骨髓病理学变化。结果K562/A02细胞分选后CD34^(+)CD38^(-)细胞比例高于分选前(P<0.05),而分选前后对多柔比星的细胞半抑制浓度差异无统计学意义(P>0.05)。A组小鼠接种10 d时肿瘤体积开始生长,13 d时出现明显增长,21 d时增长至最大。与C组相比,A组小鼠接种瘤块21 d时白细胞升高,血红蛋白和血小板减少,骨髓细胞中CD34^(+)CD117^(+)细胞比例增加(P<0.05)。接种瘤块21 d时,A组小鼠骨髓单个核细胞和移植瘤组织细胞耐药率均高于C组骨髓单个核细胞(P<0.05);并且A组小鼠肝脏、脾脏肿大,并伴有炎症坏死,股骨骨髓内可见散在原始细胞。结论采用皮下包埋方法可成功构建白血病MDR小鼠模型,且该模型具有生存情况好、干性稳定和耐药性减弱的优点,便于观察和测量移植瘤变化的优势。Objective To investigate the construction methodand its identification of mouse leukemia model with multidrug resistance(MDR).Methods The leukemia MDR cell line K562/A02 was routinely cultured.The CD34^(+)CD38^(-)cell subsets were separated by immunomagnetic bead method.The ratio of CD34^(+)CD38^(-)cells was measured by flow cytometry.The drug resistance of CD34^(+)CD38^(-)cells was analyzed by CCK^(-)8 assay.The leukemia MDR mouse model was constructed by subcutaneous embedding method(group A,10 mice),and another 10 mice injected with normal saline were taken as the controls(group C).The growth curve of tumor volume and cell count of peripheral blood were recorded.The ratio of CD34^(+)CD117^(+)cells in bone marrow cells of the mice was detected by flow cytometry.The drug resistance of xenograft cells was measured by CCK^(-)8 assay.The pathological changes of liver,spleen and bone marrow tissues were observed by HE staining.ResultsThe ratio of CD34^(+)CD38^(-)cells after the K562/A02 cells separation was higher than that before the separation(P<0.05),whereas there was no statistical difference in the IC50value for doxorubicin before and after the separation(P>0.05).The xenograft volume began to grow on the 10thday,significantly increased on the 13thday and increased to the maximum on the 21stday after the inoculation in group A.Compared with group C,the white blood cells were increased,hemoglobin and platelets were decreased,and the ratio of CD34^(+)CD117^(+)cells in bone marrow cells was increased on the 21stday after the inoculation in group A(P<0.05).The drug resistance rates of bone marrow mononuclear cells and tumor mass cells in group A were higher than those of bone marrow mononuclear cells in group C(P<0.05).The liver and spleen were oedema accompanied with inflammation and necrosis,and the scattered primordial cells were found in the bone marrow of group A.Conclusion The leukemia MDR mouse model could be successfully constructed by subcutaneous embedding method,which has the advantages of good survival,s

关 键 词：白血病 多药耐药 移植瘤 

分 类 号：R733[医药卫生—肿瘤]