Jeko-1细胞株中边群细胞的免疫表型、干细胞基因和集落形成能力的生物学特点  

Biological Characteristics of Immunophenotype,Stem Cell Gene and Colony Forming Ability in Side-Population Cells of Jeko-1 Cell Line

作　　者：周仕霞 张力 李晓明 唐君玲 无 ZHOU Shi-Xia;ZHANG Li;LI Xiao-Ming;TANG Jun-Ling;无(Department of Hematology,Affiliated Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China;Stem Cell Laboratory Affiliated Hospital of Southwest Medical Univeristy,Luzhou 646000,Sichuan Province,China;Department of Oral and Maxillofacial Surgery,Stomatological Hospital Affiliated to Southwest Medical University,Luzhou 646000,Sichuan Province,China)

机构地区：[1]西南医科大学附属医院血液内科,四川泸州646000 [2]西南医科大学附属医院干细胞实验室,四川泸州646000 [3]西南医科大学口腔医学院口腔颌面外科,四川泸州646000

出　　处：《中国实验血液学杂志》2020年第5期1558-1562,共5页Journal of Experimental Hematology

基　　金：四川省重点研发项目(2019YFS0301);四川省卫生健康委员会重点研究项目(18ZD014);西南医科大学附属医院博士科研启动基金(19032,19079);泸州市科技局应用基础研究项目(2019LZXNYDJ54)。

摘　　要：目的:初步探讨套细胞淋巴瘤中边群细胞的体外细胞生物学特性。方法:利用流式细胞术分选结合连续培养富集JeKo-1细胞中的边群细胞,并对其进行CD19、IgM免疫表型分析;用qRT-PCR进行基因验证及集落形成实验了解边群细胞及非边群细胞基因表达及体外自我更新能力的差异。结果:通过连续分选结合培养的方法可以富集JeKo-1细胞株中的边群细胞,且边群细胞中包含比例不同的4群细胞:CD19^-/IgM^-为(2.79±0.82)%,CD19^-/IgM^+为(70.99±13.61)%,CD19^+/IgM^-为(0.55±0.25)%,CD19^+/IgM^+为(25.67±14)%。Bmi-1、CD133、C-MYC、Nanog基因在边群细胞高表达,而在非边群细胞中低表达(P<0.05)。边群细胞具有更强的体外集落形成能力(P<0.05)。结论:套细胞淋巴瘤JeKo-1细胞株含有边群细胞,该群细胞可通过连续分选及培养进行富集后用于研究;JeKo-1中的边群细胞具有更强的干性基因及体外自我更新能力,并含有4种免疫表型不同的细胞群体:CD19^-/IgM^-、CD19^-/IgM^+、CD19^+/IgM^-、CD19^+/IgM^+。Objective:To investigate the in vitro biological characteristics of side-population(SP)cells in mantle cell lymphoma(MCL).Methods:The SP cells in JeKo-1 cells were enriched by continuous culture and sorted by flow cytometry(FCM),and the expression of CD19 and IgM were analyzed.The differences of gene expression and their self-renewal ability between SP cells and non-SP cells were estimated by qRT-PCR and colony-forming assay(CFA)respectively.Results:SP cells in JeKo-1 cells could be enriched by continuous sorting and in vitro cultureing,and it was found that CD19^-/IgM^-accounted for(2.79±0.82)%,CD19^-/IgM^+accounted for(70.99±13.61)%,CD19^+/IgM^-accounted for(0.55±0.25)%,and CD19^+/IgM^+accounted for(25.67±14)%.Bmi-1,CD133,C-MYC and Nanog genes were highly expressed in SP cells,but were low expressed in non-SP cells(P<0.05).Compared with non-SP cells,SP cells had stronger colony forming ability(P<0.05).Conclusion:JeKo-1 cell line of mantle cell lymphoma contains SP cells,which can be enriched by continuous sorting and culture for research.The SP cells in JeKo-1 contain four different cell populations with different immunophenotypes:CD19^-/IgM^-、CD19^-/IgM^+、CD19^+/IgM^-、CD19^+/IgM^+,and show stronger gene expression of stem cells and self-renewal ability in vitro.

关 键 词：JeKo-1细胞 套细胞淋巴瘤 边群细胞 干细胞基因 

分 类 号：R733.1[医药卫生—肿瘤]