miR-451在白血病细胞株K562/A02多药耐药中的作用及相关机制  

The Role and Mechanism of MiR-451 in Multidrug Resistance of Leukemia Cell Line K562/A02

作　　者：冯延丽 苏保雄[1] 葛繁梅[1] 戴崇文[2] FENG Yan-Li;SU Bao-Xiong;GE Fan-Mei;Dai Chong-Wen(Department of Hematology.Yan'an University Affiliated Hospital,Yan'an 716000,Shaanxi Province,China;Depariment of Hematology,The Second Xiangya Hospital of Central South University,Changsha 410011,Hunan Province,China)

机构地区：[1]延安大学附属医院血液科,陕西延安716000 [2]中南大学湘雅二医院血液科,湖南长沙410011

出　　处：《中国实验血液学杂志》2023年第3期685-692,共8页Journal of Experimental Hematology

摘　　要：目的:检测miR-451、ABCB1、ABCC2在白血病药物敏感细胞株K562及其耐药株K562/A02中的差异表达,探讨miR-451与ABCB1、ABCC2表达的调控关系及miR-451参与白血病耐药的相关机制。方法:CCK-8法检测K562/A02及K562细胞的耐药性,实时荧光定量PCR(qRT-PCR)验证miR-451在K562及K562/A02细胞中的差异表达,将miR-451模拟物(mimic)及其阴性对照(miR-NC)、miR-451抑制剂(inhibitor)及其阴性对照(miR-inNC)分别转染K562及K562/A02细胞,应用q RT-PCR、Western blot检测ABCB1、ABCC2在K562、K562/A02细胞及转染后各组细胞中的mRNA、蛋白表达水平。结果:K562/A02细胞对阿霉素的耐药是其亲本细胞系K562的177倍。与K562细胞相比,K562/A02细胞中miR-451明显高表达(P<0.001),ABCB1、ABCC2在K562/A02中的mRNA及蛋白表达水平均显著高于K562细胞(P<0.001)。K562/A02细胞转染miR-451 inhibitor后,miR-451表达显著下调(P<0.001),对化疗药物的敏感性显著增强(P<0.05),ABCB1、ABCC2 mRNA及蛋白表达水平显著降低(P<0.01)。K562细胞转染miR-451 mimic后,miR-451表达显著上调(P<0.001),ABCB1、ABCC2 mRNA及蛋白表达水平显著增高(P<0.01)。结论:miR-451、ABCB1及ABCC2在白血病敏感细胞株K562及其耐药株K562/A02中的表达存在显著差异,miR-451可能通过调控ABCB1、ABCC2的表达对白血病细胞的耐药性产生影响。Objective:To detect the differential expressions of miR-,ABCB1 and ABCC2 in drug-sensitive leukemia cell line K562 and drug-resistant cell line K562/A02,and explore the regulatory relationship between miR-451 and the expressions of ABCB1 and ABCC2,and the mechanism of miR-451 involved in drug resistance in leukemia.Methods:CCK-8 assay was used to detect the drug resistance of K562/A02 and K562 cells.Quantitative Real-time PCR(qRT-PCR)was used to verify the differential expressions of miR-451 in K562 and K562/A02 cells.MiR-451 mimic and negative control(miR-NC),miR-451 inhibitor and negative control(miR-inNC)were transfected into K562 and K562/A02 cells respectively,then qRT-PCR and Western blot were used to detect the expression levels of mRNA and protein of ABCB1 and ABCC2 in K562 and K562/A02 cells and the transfected groups.Results:The drug resistance of K562/A02cells to adriamycin was 177 times higher than that of its parent cell line K562.Compared with K562 cells,the expression of miR-451 in K562/A02 cells was significantly higher(P<0.001),and the mRNA and protein expression levels of ABCB1 and ABCC2 in K562/A02 cells were significantly higher than those in K562 cells(P<0.001).After transfected with miR-451 inhibitor,the expression of miR-451 was significantly down-regulated in K562/A02 cells(P<0.001),the sensitivity to chemotherapy drugs was significantly enhanced(P<0.05),and the mRNA and protein expressions of ABCB1and ABCC2 were significantly decreased(P<0.01).After transfected with miR-451 mimic,the expression of miR-451 was significantly upregulated in K562 cells(P<0.001),and the mRNA and protein expressions of ABCB1 and ABCC2 were significantly increased(P<0.01).Conclusion:There are significant differences in the expressions of miR-451,ABCB1and ABCC2 between the drug-sensitive leukemia cell line K562 and drug-resistant cell line K562/A02,which suggests that mi R-451 may affect the drug resistance of leukemia cells by regulating the expression of ABCB1 and ABCC2.

关 键 词：白血病 多药耐药 miR-451 ABCB1 ABCC2 

分 类 号：R733.7[医药卫生—肿瘤]