RNA干扰YB-1表达对阿霉素诱导性耐药的K562细胞mdr1基因表达的影响  

Effect of regulation of Y-box protein 1 by RNA interference on the doxorubicin induced mdrl gene expression in K562 cells

作　　者：沈慧玲[1] 许文林[1] 陈巧云[1] 王法春[1] 费霞[1] 

机构地区：[1]江苏大学附属人民医院中心实验室,镇江212002

出　　处：《中华血液学杂志》2011年第6期383-387,共5页Chinese Journal of Hematology

基　　金：江苏省卫生重大项目（K2005017）;江苏省医学重点人才资助项目（RC2007034）

摘　　要：目的探讨Y-盒结合蛋白1（YB-1）对白血病细胞系K562细胞药物诱导性mdrl基因表达的调控作用。方法阿霉素间歇性长期作用于K562细胞，剂量逐渐增加，诱导细胞耐药。RT-PCR检测mdrl和YB-1基因表达，流式细胞术检测mdrl基因编码的P-糖蛋白（P-gP）表达，蛋白质印迹法检测YB-1蛋白核易位程度。通过RNA干扰技术使K562细胞中YB-1基因表达沉默，再用阿霉素处理YB-1基因沉默细胞，诱导细胞耐药，用RT-PCR、流式细胞术分别检测mdrlmRNA和P-gp的表达。结果经阿霉素作用后K562细胞mdrl基因转录上调，P—gP表达增加，同时YB-1基因转录也增强，且出现明显的核易位。YB-1基因沉默后，mdrl基因诱导性表达明显减少，阿霉素浓度为0．03、0．05μg／ml时 mdr1 mRNA表达水平分别较未行shRNA干扰组降低（53．7±0．1）％和（64．3±0．0）％，P-gP表达呈同样趋势。结论阿霉素诱导K562细胞耐药形成的过程中，YB-1对于mdr1基因的转录活化起一定的作用。Objective To investigate the effect of YB-I on the transcription of induced mdrl gene expression in K562 cells. Methods K562 cells were treated with doxorubicin （DOX） at different concentrations and times. Expression of mdrl and YB-1 genes was examined by RT-PCR and P-glycoprotein （P-gp） by flow cytometry. Cyto/nuclear protein was extracted for YB-1 detection by Western blotting. The expression of YB-1 gene in K562 cells was inhibited by YB-1 gene specific RNA interference （RNAi） , then the expression of mdrl and P-gp in YB-1 gene silenced cells treated with DOX was detected. Results The mdrl gene as well as its cmTesponding protein P-gp was highly expressed in DOX exposed K562 cells. DOX up-regulated the expression of YB-I gene, and promoted YB-1 protein nuclear translocation. On YB-1 gene silenced, the expressions of mdrl gene and P-gp were obviously down-regulated in DOX treated K562 cells. Conclusion Doxorubicin can induce the expression of mdrl gene in 1（562 cells, which may result from the transcription of mdrl gene by activated YB-1.

关 键 词：Y-盒结合蛋白1 基因 mdr1 多药耐药 转录调节 

分 类 号：R730.2[医药卫生—肿瘤]