不同多发性骨髓瘤细胞系移植小鼠的成瘤情况分析  

Analysis of Tumorigenesis in Mice Transplanted with Different Multiple Myeloma Cell Lines

作　　者：安娜 刘加军[1] AN Na;LIU Jia-Jun(Department of Hematology,The Third Affiliated Hospital of Zhongs han University,Guangzhou 510630,Guangdong Province,China;Department of Hematology,Shenzhen Second People's Hospital,Shenzhen 518035,Guangdong Province,China)

机构地区：[1]中山大学附属第三医院血液科,广东广州510630 [2]深圳市第二人民医院血液科,广东深圳518035

出　　处：《中国实验血液学杂志》2019年第5期1522-1529,共8页Journal of Experimental Hematology

基　　金：中国博士后金(2017M612815);深圳市科创委基础研究与自由探索项目(JCYJ20170306092132534)

摘　　要：目的:探讨在无γ射线照射条件下,几种常见的多发性骨髓瘤(MM)细胞系移植小鼠的成瘤情况,以及构建MM疾病模型便于体内实验研究。方法:利用MM细胞系LP-1、OPM2、RPMI 8226和MOLP8,以及通过转染带有Luciferase荧光素酶的慢病毒载体得到的RPMI-Luc-Puro(载体带Puromycin抗性基因)、RPMI-LucmCherry(载体带mCherry标记基因)和MOLP8-Luc-Puro稳定细胞系,经皮下或者尾静脉植入NOD/SCID或NSG小鼠。观察其肿瘤生长,测量肿瘤大小;应用流式细胞术检测小鼠尾血中CD138^+肿瘤细胞的比例;应用血清免疫固定电泳检测外周血中的游离轻链;免疫组织化学染色法鉴定肿瘤类型;应用活体成像技术监测全身肿瘤信号。结果:LP-1和OPM2细胞皮下植入NOD/SCID小鼠各21只,观察至7周均未见成瘤。RPMI 8226细胞皮下植入的NOD/SCID小鼠15只,1周后可观察到肿瘤形成,截至7周时,成瘤率80%(12/15)。血清免疫固定电泳可以检测到λ轻链,尾血中未检测到CD138^+的肿瘤细胞;免疫组织化学染色支持浆细胞肿瘤。RPMI-Luc-Puro、RPMI-Luc-mCherry和MOLP8-Luc-Puro细胞皮下植入NOD/SCID小鼠各2只。活体成像显示RPMI-Luc-mCherry细胞植入小鼠无明显肿瘤信号,RPMI-Luc-Puro和MOLP8-Luc-Puro细胞植入小鼠1周时能探测到明显肿瘤信号,但前者2周时信号消失,后者观察至3周时肿瘤信号仍存在;这两种细胞皮下植入NSG小鼠时,两者的肿瘤信号均能持续存在。RPMI 8226、RPMI-Luc-Puro、RPMI-Luc-mCherry和MOLP8-Luc-Puro细胞经尾静脉植入NOD/SCID或NSG小鼠均未出现全身播散的肿瘤信号,仅MOLP8-Luc-Puro细胞植入的1只NSG小鼠出现过短暂的全身播散信号。结论:在无γ射线照射条件下,MM细胞系植入小鼠可以成瘤,有局部成瘤倾向,全身播散能力低;不同细胞系成瘤性存在较大差异,载体改造会降低细胞的成瘤能力;免疫缺陷更严重的NSG小鼠有利于肿瘤生长。Objective:To investigate the tumorigenicity of several multiple myeloma(MM)cell lines transplanted in mice withoutγ-ray irradiation and to construct the MM disease model to facilitate in vivo experiments.Methods:NOD/SCID or NSG mice were subcutaneously or caudally transplanted with MM cell lines(LP-1,OPM2,RPMI 8226 and MOLP8),or cell lines with luciferase(RPMI-Luc-Puro,RPMI-Luc-m Cherry and MOLP8-Luc-Puro).Tumor growth was observed by measuring the tumor size with a caliper.CD138^+tumor cells in peripheral blood were detected by flow cytometry.The free light chain in mouse serum was detected by immunofixation electrophoresis.Tumor type was identified by immunohistochemistry.Results:Twenty one NOD/SCID mice were subcutaneously transplanted with LP-1 cells or OPM2 cells respectively,and no tumors formed till 7 weeks after transplantation.Fifteen NOD/SCID mice subcutaneously transplanted with RPMI 8226 cells showed tumor formation one week later.As of 7 weeks,the rate of tumorigenesis was 80%(12/15).Serumλlight chain was detected and no CD138^+tumor cells were detected in peripheral blood.Two NOD/SCID mice each were subcutaneously transplanted with RPMI-Luc-Puro,RPMI-Luc-mcherry and MOLP8-Luc-Puro cells respectively.No tumor signal was detected through IVIS in RPMI-Luc-mcherry cells-transplanted mice.There was tumor signal at 1 week in RPMI-Luc-Puro and MOLP8-Luc-Puro cells-transplanted mice,the former disappeared at 2 weeks and the latter persisted more than 3 weeks.NSG mice subcutaneously transplanted with both cells persistently displayed the tumor signal.Neither NOD/SCID nor NSG mice transplanted with RPMI 8226,RPMI-Luc-Puro,RPMI-Luc-mcherry or MOLP8-Luc-Puro cells through tail vein developed the tumor signal.Only one NSG mice transplanted with MOLP8-Luc-Puro cells appeared transient tumor signal.Conclusion:Unirradiated mice transplanted with MM cell lines tended to develop local tumor,and failed to develop disseminated tumor.The tumorigenicity of different cell lines is quite different and the vector transfec

关 键 词：多发性骨髓瘤 NOD/SCID小鼠 多发性骨髓瘤细胞移植 动物模型 

分 类 号：R733.3[医药卫生—肿瘤]