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浙江大学学报(医学版)  2011, Vol. 40 Issue (5): 489-494    DOI: 10.3785/j.issn.1008-9292.2011.05.005
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细胞因子诱导K562/MDR1分化为具有多药耐药特征的树突状细胞的实验研究
盛立霞1,2,谢晓宝3,欧阳桂芳2,王怡2,朱慧玲2,黄河1
1.浙江大学医学院附属第一医院骨髓移植中心,浙江 杭州 310003; 2.宁波市第一医院血液科,浙江 宁波 315010; 3.苏州大学附属第三医院血液科,江苏 常州 213003
Induction of dendritic cells with multidrug resistance from K562/MDR1 cells

SHENG Li-xia 1,2, XIE Xiao-bao 3, OU-YANG Gui-fang 2 , WANG Yi 2, ZHU Hui-ling 2, HUANG He1
1.The First Affiliated Hospital,Zhejiang University School of Medicine,Hangzhou 310003,China; 2.The First Ningbo Municipal Hospital,Ningbo 315007,China; 3.The Third Affiliated Hospital of Soochow University,Changzhou 213003,China
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摘要: 目的:研究多药耐药基因1(multidrug resistance gene1,MDR1)转染的K562细胞是否能够在细胞因子诱导下向树突状细胞(DC)诱导分化,获得多药耐药特性的K562来源的DC。
方法:将K562/MDR1和K562细胞分别用GM-CSF+IL-4诱导培养DC,并以TNF-α促进DC成熟,于第14天收获细胞K562/MDR1-DC及K562-DC。通过流式细胞仪(FCM)检测细胞分化前后的CD1a、CD83、CD80、CD86、HLA-ABC和HLA-DR的表达;异基因混合淋巴细胞反应(Allo-MLR)检测其抗原递呈功能;流式细胞仪检测Pgp的表达及细胞内柔红霉素(DNR)的蓄积浓度;MTT法测定K562/MDR1-DC及K562-DC对化疗药物长春新碱(VCR)、阿霉素(ADM)的IC50。
结果:K562/MDR1和K562细胞均能在适当细胞因子组合下分化为具有DC形态特征和表型特征的K562/MDR1-DC及K562-DC,K562/MDR1-DC在Allo-MLR反应中显示出比K562-DC更强的抗原递呈功能。与K562-DC相比,K562/MDR1-DC具有膜Pgp分子的高表达和对胞内DNR的外排作用,对化疗药物VCR、ADM有较高的IC50。
结论:转染MDR1基因不影响K562向树突状细胞分化的能力;并且K562/MDR1-DC具有Pgp的高表达,获得多药耐药特征。
关键词: 树突细胞 细胞因子类 P糖蛋白 K562细胞 柔红霉素/药理学 基因 转染    
Abstract: Objective: To induce the differentiation of K562/MDR1 cells into dendritic cells (DC) with multidrug resistance property.
Methods: K562/MDR1 cells and K562 cells were cultured in the presence of GM-CSF and IL-4 to generate DC and matured by TNF-α.On d14 K562/MDR1-DC and K562-DC cells were harvested and the expressions of CD1a,CD83,CD80,CD86,HLA-ABC and HLA-DR were assessed by flow cytometry (FCM).The antigen presentation function of K562/MDR1-DC and K562-DC was determined by allogenic mixed lymphocyte reaction (Allo-MLR).The expression of P-glycoprotein and the intracellular accumulation of daunorubicin (DNR) were detected by FCM.The sensitivity of K562/MDR1-DC and K562-DC cell to vincristine,adriamycin was measured using MTT assay.
Results: Both K562/MDR1 and K562 cells were differentiated into dendritic cells in the presence of cytokine cocktails,showing the morphologic and immunophenotypic characteristics of DC.K562/MDR1-DC more markedly enhanced proliferation of allogenetic lymphocytes in MLR than K562-DC.High level expression of P-glycoprotein and efflux of DNR were demonstrated in K562/MDR1-DC.K562/MDR1-DC showed multidrug resistance property,with higher IC50 to VCR and ADM than that of K562-DCs.
Conclusion: K562/MDR1 cells can be differentiated into DC with the presence of cytokines,the induced K562/MDR1-DC cells express high level of P-glycoprotein and acquire the multidrug resistance property.
Key words: Dendritic cells    Cytokines    P-glycoprotein    K562 cells    Daunorubicin/pharmacol    Genes    Transfection
出版日期: 2011-09-25
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引用本文:

盛立霞;谢晓宝;欧阳桂芳;王怡;朱慧玲;黄河. 细胞因子诱导K562/MDR1分化为具有多药耐药特征的树突状细胞的实验研究[J]. 浙江大学学报(医学版), 2011, 40(5): 489-494.

链接本文:

https://www.zjujournals.com/med/CN/Y2011/V40/I5/489

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