牛蒡子苷元对白血病K562/A02细胞耐药性的逆转作用及机制  

Reversal Effect of Arctigenin on the Drug Resistance in Leukemia K562/A02 Cells and Its Mechanism

作　　者：邹琳 方烨 何威 ZOU Lin;FANG Ye;HE Wei(General Department,General Hospital of Central Theater Command of the Chinese People′s Liberation Army,Wuhan 430070,Hubei Province,China)

机构地区：[1]中国人民解放军中部战区总医院综合科,湖北武汉430070

出　　处：《中国实验血液学杂志》2024年第2期409-415,共7页Journal of Experimental Hematology

摘　　要：目的:研究牛蒡子苷元对白血病耐药细胞株K562/A02阿霉素耐药性的影响及其作用机制。方法:体外培养人白血病细胞株K562及阿霉素耐药细胞株K562/A02,使用2.5-50μmol/L阿霉素处理,CCK-8检测细胞生长情况并计算药物半数抑制浓度(IC_(50))。采用不同浓度的牛蒡子苷元(1、2、4、8、16 mmol/L)处理K562/A02细胞,检测牛蒡子苷元对K562/A02细胞的影响,筛选适用浓度用于后续实验。在2 mmol/L牛蒡子苷元处理的K562/A02细胞中加入5μmol/L阿霉素,流式细胞术检测细胞凋亡,Western blot检测P-gp、MRP、cleaved caspase-3、Bax、Bcl-2以及TLR4/NF-κB信号通路蛋白的表达。在牛蒡子苷元和阿霉素共处理的K562/A02细胞中转染TLR4过表达质粒,检测细胞的药物敏感性和凋亡水平。结果:阿霉素对K562/A02细胞的IC_(50)为36.57μmol/L,高于K562细胞(1.30μmol/L)。当牛蒡子苷元浓度≤2 mmol/L时,K562/A02的细胞生长不会受到明显抑制。经2 mmol/L的牛蒡子苷元处理后,阿霉素对K562/A02细胞的IC_(50)明显降低。与对照组相比,牛蒡子苷元组K562/A02细胞凋亡率明显上升,cleaved caspase-3、Bax蛋白的表达明显上调,P-gp、MRP、Bcl-2、TLR4、My D88和p-NF-κB蛋白的表达明显下调,差异均具有统计学意义(P<0.05)。在转染TLR4过表达质粒后,牛蒡子苷元处理的K562/A02细胞对阿霉素的敏感性明显降低(P<0.05),细胞凋亡下降,并显著提升了P-gp、MRP、Bcl-2和TLR4/NF-κB信号通路蛋白的表达(P<0.05),同时降低了cleaved caspase-3和Bax蛋白的表达(P<0.05)。结论:牛蒡子苷元可能通过抑制TLR4/NF-κB信号通路从而逆转人白血病耐药细胞株K562/A02对阿霉素的耐药性。Objective:To study the effect of arctigenin(ARG)on adriamycin(ADM)resistance of leukemia cell line K562/A02 and the underlying mechanism.Methods:Human leukemia cell line K562 and ADM-resistant cell line K562/A02 were cultured and treated with 2.5-50µmol/L ADM.Cell proliferation was measured using CCK-8 method,and half maximal inhibitory concentration(IC_(50))was calculated.K562/A02 cells were treated with different concentrations of ARG(1,2,4,8,16 mmol/L)to detect the effect of ARG on K562/A02 cells,and a suitable concentration(2 mmol/L)was selected for subsequent experiments.K562/A02 cells were treated with 2 mmol/L ARG and 5µmol/L ADM,and cell apoptosis was detected by flow cytometry,the expression of P-gp,MRP,cleaved caspase-3,Bax,Bcl-2 proteins and the TLR4/NF-κB signaling pathway-related proteins were measured by Western blot.TLR4 overexpression plasmid was transfected into K562/A02 cells which were co-treated with ARG and ADM,then drug sensitivity and cell apoptosis were measured.Results:The IC_(50)value of ADM on K562/A02 cells was 36.57µmol/L,which was significantly higher than that on K562 cells(1.30µmol/L).ARG with a concentration of≤2 mmol/L did not have a significant effect on K562/A02 cells.2 mmol/L ARG significantly reduced the IC_(50)of ADM on K562/A02 cells.In 5µmol/L ADM-treated K562/A02 cells,compared with the control group,the apoptosis rate of K562/A02 cells in the ARG group was significantly increased,the expressions of cleaved caspase-3,Bax proteins were significantly upregulated,the expressions of P-gp,MRP,Bcl-2,TLR4,MyD88,and p-NF-κB proteins were significantly downregulated,and the differences were statistically significant(P<0.05).After transfection with TLR4 overexpression plasmid,the sensitivity of ARG-treated K562/A02 cells to ADM was reduced(P<0.05),the cell apoptosis was decreased,and the expressions of P-gp,MRP,Bcl-2 and TLR4/NF-κB signaling pathway-related proteins were significantly elevated,while the expressions of cleaved caspase-3 and Bax proteins were significantly

关 键 词：牛蒡子苷元 白血病 K562/A02细胞 耐药 TLR4/NF-κB信号通路 

分 类 号：R733.7[医药卫生—肿瘤]