白血病耐药细胞中多药耐药基因MDR1与核转录因子κB的关系  

Relationship between multi-drug resistance gene MDR1 and NF-kappa B in human drug-resistance erythroleukemia cells

作　　者：杨冬艳[1] 张宏宇[2] 孔晓霞[2] 孙连坤[2] 史纪文[1] 

机构地区：[1]吉林大学中日联谊医院超声科,吉林长春130033 [2]吉林大学基础医学院病理生理学教研室,吉林长春130021

出　　处：《吉林大学学报（医学版）》2009年第4期650-655,共6页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科技发展计划项目资助课题(200705373)

摘　　要：目的:研究核转录因子κB与多药耐药基因MDR1在白血病化疗耐药中的作用,阐明核转录因子κB(NF-κB)与白血病多药耐药的关系及相关分子机制。方法:白血病细胞K562和阿霉素(ADM)诱导的耐药细胞K562/ADM分别应用ADM(0、0.25、0.50、1.00和2.00μg.L-1)或丝裂霉素(MIT)(0、2、4、8和16 mg.L-1)作用48 h,通过MTT法检测细胞生存率,计算细胞耐药指数;RT-PCR方法检测MDR1和IκBmRNA表达;Western blotting方法检测P-gp和P65的蛋白表达。结果:ADM和MIT分别作用后,与空白对照组比较,白血病细胞K562细胞生存率下降(P<0.01),呈剂量依赖性;ADM和MIT对耐药细胞K562/ADM生存率作用不明显;在mRNA水平上,与K562细胞比较,K562/ADM细胞的MDR1 mRNA高表达(P<0.01)。与空白对照组比较,ADM 0.50和1.00μg.L-1或MIT 4和8 mg.L-1分别作用K562细胞和K562/ADM细胞,IκBαmRNA表达均下降(P<0.01),并呈剂量依赖性;在蛋白水平上,与K562细胞比较,K562/ADM细胞的P-gp蛋白高表达(P<0.01)。与空白对照组比较,ADM 0.50和1.00μg.L-1或MIT 4和8 mg.L-1分别作用K562/ADM细胞,P-gp蛋白表达量升高(P<0.01),同时P65蛋白表达量升高,且呈药物剂量依赖性(P<0.01)。结论:MDR1基因及P-gp蛋白的高表达很可能是白血病细胞K562多药耐药性形成的关键机制,而NF-κB的激活可能参与了多药耐药的调控。Objective To observe the effect of nuclear factor kappa B（NF-κB） and multi-drug resistance gene MDR1 on leukemia chemotherapy resistance,to demonstrate the relationship between NF-κB and multidrug-resistance and the related mechanism.Methods Erythroleukemia cells K562 and adriamycin（ADM）-resistance cells K562/ADM were treated with ADM（0,0.25,0.50,1.00,2.00 μg·L^-1 or mitomycin（MIT）（0,2,4,8,16 mg·L^-1 for 48 h. The cell viability was detected by MTT assay,the cell resistant factor was calculated,MDR1 and IκB mRNA levels were detected by RT-PCR;Pgp and P65 protein levels were detected by Western blotting.Results Compared with control group,0.50 and 1.00 μg·L^-1ADM or 4 and 8 mg·L^-1MIT inhibited the cell viability significantly in erythroleukemia cells K562 in a dose-dependent manner（P〈0.01）;but had no significant effect on drug-resistance cells;on the mRNA level,compared with K562 cells,the expression level of MDR1 mRNA in K562/ADM cells was higher（P〈0.01）.Compared with control group,the expressions of IκBα mRNA decreased（P〈0.01） in K562 cells or K562/ADM cells treated with 0.50 and 1.00 μg·L^-1ADM or 4 and 8 mg·L^-1MIT respectively in a dose-dependent manner.On the protein level,compared with K562 cells,the expression of P-gp protein increased（P〈0.01）;compared with control group,the expression of P-gp protein increased（P〈0.01）,the expression of P65 protein increased at the same time in a dose-dependent manner in K562/ADM cells treated with 0.50 and 1.00 μg·L^-1ADM or 4 and 8 mg·L^-1MIT（P〈0.01）.Conclusion The high expression of MDR1 gene and P-gp protein may be the essential mechanism of drug resistance in K562 cells,and the activation of NF-κB may be involved in the regulation of multi-drug resistance.

关 键 词：多药耐药基因 核转录因子ΚB P65 

分 类 号：R730.53[医药卫生—肿瘤]