Sheng Li-xia, Xie Xiao-bao, Ou-Yang Gui-fang, Wang Yi, Zhu Hui-ling, Huang He
The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2011 Sep;40(5):489-94. doi: 10.3785/j.issn.1008-9292.2011.05.005.
To induce the differentiation of K562/MDR1 cells into dendritic cells (DC) with multidrug resistance property.
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.
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 allogeneic 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 IC(50) to VCR and ADM than that of K562-DCs.
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.
诱导具有多药耐药特性的K562/MDR1细胞分化为树突状细胞(DC)。
将K562/MDR1细胞和K562细胞在GM-CSF和IL-4存在的条件下培养以生成DC,并通过TNF-α使其成熟。在第14天收获K562/MDR1-DC和K562-DC细胞,采用流式细胞术(FCM)评估CD1a、CD83、CD80、CD86、HLA-ABC和HLA-DR的表达。通过异体混合淋巴细胞反应(Allo-MLR)测定K562/MDR1-DC和K562-DC的抗原呈递功能。采用FCM检测P-糖蛋白的表达及柔红霉素(DNR)的细胞内蓄积。使用MTT法测定K562/MDR1-DC和K562-DC细胞对长春新碱、阿霉素的敏感性。
在细胞因子混合物存在的情况下,K562/MDR1和K562细胞均分化为树突状细胞,表现出DC的形态学和免疫表型特征。与K562-DC相比,K562/MDR1-DC在MLR中更显著地增强了异体淋巴细胞的增殖。在K562/MDR1-DC中证实了P-糖蛋白的高水平表达和DNR的外排。K562/MDR1-DC表现出多药耐药特性,对VCR和ADM的IC(50)高于K562-DC。
在细胞因子存在的情况下,K562/MDR1细胞可分化为DC,诱导产生的K562/MDR1-DC细胞表达高水平的P-糖蛋白并获得多药耐药特性。