荧光素酶标记的KG1a细胞构建急性髓系白血病小鼠模型及其鉴定  

The Establishment and Identification of Acute Myeloid Leukemia NOD-SCID-IL2rg^(-/-)Mice Model by Using Luciferase-Expressing KGla Cells

作　　者：张维娅 曾高淳 陈晓梅[2] 耿素霞[2] 王玉连 罗琼[2] 罗柳萍[2] 赖沛龙[2] 翁建宇[2] 杜欣 ZHANG Wei-Ya;ZENG Gao-Chun;CEHN Xiao-Mei;GENG Su-Xia;WANG Yu-Lian;LUO Qiong;LUO Liu-Ping;LAI Pei-Long;WENG Jian-Yu;DU Xin(School of Medicine,South China University of Technology,Guangzhou 510006,Guangdong Province,China;Department of Hematology,Guangdong Provincial People's Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510080,Guangdong Province,China)

机构地区：[1]华南理工大学医学院,广东广州510006 [2]广东省人民医院(广东省医学科学院)血液科,广东广州510080

出　　处：《中国实验血液学杂志》2021年第5期1429-1435,共7页Journal of Experimental Hematology

基　　金：国家自然科学基金(81870121);广东省自然科学基金(2019A1515012049,2019A1515010094)。

摘　　要：目的:利用荧光素酶标记的KG1a急性髓系细胞株构建能够在活体水平观察肿瘤负荷的急性髓系白血病小鼠模型。方法:经慢病毒及嘌呤霉素筛选构建能稳定表达荧光素酶的KG1a细胞(KG1a-Luc细胞);将18只8-12周龄的雄性NOD-SCID-IL2rg^(-/-)小鼠随机均分为2组,模型组小鼠每只尾静脉注射200μl含有5×10^(6)KG1a-Luc细胞的PBS,对照组小鼠每只尾静脉注射200μl PBS;通过生物发光成像技术于活体水平监测小鼠肿瘤负荷;尾静脉采血后,经流式细胞术检测小鼠外周血中肿瘤细胞占比;颈椎脱臼法处死小鼠后,取小鼠骨髓及脾脏制备涂片,取肝脏制备石蜡切片,染色后镜下观察肿瘤浸润情况;记录并比较2组小鼠生存时间。结果:经过慢病毒转染、嘌呤霉素筛选后能够得到稳定表达荧光素酶的KG1a细胞;d 17使用生物发光成像技术显示模型组小鼠出现明显荧光;各模型组小鼠的外周血中均能检出KG1a-Luc肿瘤细胞群,平均比率为(16.27±6.66)%;形态学及病理学检查显示,骨髓、脾脏及肝脏均有KG1a-Luc细胞浸润;与对照组相比,模型组小鼠的生存时间显著缩短,其中位生存时间为30.5 d(95%CI:0.008-0.260)。结论:NOD-SCID-IL2rg-/-小鼠经尾静脉注射5×10^(6)KG1a-Luc细胞后能够成功稳定建立在活体水平观察肿瘤负荷急性髓系白血病小鼠模型。Objective:To establish the in vivo traceable acute myeloid leukemia mice model with LuciferaseExpressing KGla Cells.Methods:KGla cells with stable luciferase gene expression(called as KG1α-Luc cells)were constructed by lentivirus transfection,then sifted out by puromycin.Eighteen male NOD-SCID-IL2 rg^(-/-)mice aged 8 to12 weeks were randomly and equally divided into two groups:the control group and the KGla-Luc group.The mice in KG1 a-Luc group were injected with 200μl PBS containing 5 x10^(6)KGla-Luc cells through tail veins,and the mice in control group were injected with 200μl PBS only.The bioluminescence imaging technology was used to monitor the tumor burden in vivo.The peripheral blood of the mice in both groups was analyzed by flow cytometry.After the mice were sacrificed,there were pathologic evaluations:bone marrow and spleens made into smears,and livers sliced to get paraffin sections.The survival time of the mice in the two groups was recorded and compared.Results:KGla cells expressing luciferase stably were successfully obtained.The tumor luminescence wildly spread at day 17 captured by in vivo imaging.The KG1 a-Luc tumor cells could be detected in the peripheral blood of the mice,with the average percentage of(16.27±6.66)%.The morphology and pathology result showed that KG1 a-Luc cells infiltrate was detected in bone marrow,spleens and livers.The survival time of the KG1 a-Luc mice was notably shorter as compared with those in the control group,the median survival time was 30.5 days(95%CI:0.008-0.260).Conclusion:The acute myeloid leukemia NOD-SCID-IL2 rg^(-/-)mouse model was successfully established by tail vein injection of 5×10^(6)KG1 a-Luc cells.

关 键 词：急性髓系白血病 KG1a细胞 荧光素酶 动物模型 活体荧光成像 

分 类 号：R733.71[医药卫生—肿瘤]