ZNRD1小干扰RNA对HL-60/VCR细胞耐药性逆转的研究  

Reversal of multidrug resistance of HL-60/VCR cells by down-regulation of ZNRD1 with ZNRD1 siRNA

作　　者：朴瑛[1] 刘丽梅[1] 洪流[2] 周凡[1] 刘彦琴[1] 刘景华[1] 白颖[1] 

机构地区：[1]沈阳军区总医院血液内科,沈阳110016 [2]第四军医大学西京医院消化内科,西安710032

出　　处：《肿瘤》2006年第4期339-342,共4页Tumor

基　　金：沈阳军区总医院青年启动基金资助项目(编号:03YC06)

摘　　要：目的:探讨转录相关锌带蛋白基因(zincribbondomaincontaining1protein,ZNRD1)作为靶分子进行白血病多药耐药基因治疗的可行性。方法:Westernblot检测ZNRD1在白血病细胞中的表达;构建ZNRD1基因的小干扰RNA载体并将其转导入HL60/VCR细胞,G418筛选稳定转染细胞株;MTT法检测细胞转染前后生长速度和药物敏感性的变化;Westernblot检测细胞转染前后凋亡相关蛋白bcl2、Bax和耐药相关蛋白Pgp、MRP的表达变化。结果:ZNRD1在HL60/VCR细胞中的表达明显高于HL60细胞;成功建立了低表达ZNRD1的白血病细胞模型;转染后的细胞对长春新碱和阿霉素的敏感性增强,且低表达Pgp和bcl2。结论:该小干扰RNA真核表达载体能在一定程度上逆转了白血病细胞的耐药性。Objective： To evaluate the feasibility of gene therapy using Zinc ribbon domaincontaining 1 （ZNRD1） as target for multiple drug resistance of leukemia. Methods： The expression of ZNRD1 in leukemia cells was detected by Western blot. The expression vectors of human ZNRD1 small interfering RNA （siRNA） were constructed and transfected into HL 60/VCR cells. Stable transfeetants were obtained by G418 screening. The growth of leukemia cells was measured by using MTT assay. The expression of Bel-2,Bax,P-gp, and multidrug resistance-associated protein （MRP） was analyzed by Western blot. Results： The expression of ZNRD1 was significantly higher in HL-60/VCR cells than that in HL-60 cells. After transfection with mU6 pro-ZNRD1 siRNA,ZNRD1 expression decreased dramatically in leukemia HL-60/VCR cells. The IC50 value of transfected cells to vincristine and adriamycin was significantly reduced as compared with that of the control. The expression of Bcl.-2 and P-gp in transfected cells decreased as compared with that of the control, while the expression of Bax and MRP had no significant change. Conclusiom ZNRD1 siRNA constructs partly reversed the multiple drug resistance of HL-60/VCR cells to conventional chemotherapeutic agents,

关 键 词：白血病 实验性 抗药性 多药 RNA干扰 基因 ZNRD1 

分 类 号：R733.7[医药卫生—肿瘤] Q784[医药卫生—临床医学]