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  

Induction of dendritic cells with multidrug resistance from K562/MDR1 cells

作　　者：盛立霞[1,2] 谢晓宝[3] 欧阳桂芳[2] 王怡[2] 朱慧玲[2] 黄河[1] 

机构地区：[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

出　　处：《浙江大学学报（医学版）》2011年第5期489-494,共6页Journal of Zhejiang University(Medical Sciences)

基　　金：宁波市自然科学基金资助项目(2009C50016)

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。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.

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 

分 类 号：R733.7[医药卫生—肿瘤]