柔红霉素对HL-60敏感细胞miRNA-21等5种miRNA表达的影响  

The effects of daunorubicin on the expression of miRNA in HL-60 cells

作　　者：燕华[1] 李卫[2] 

机构地区：[1]河南省郑州市儿童医院血液科,450053 [2]广西医科大学科学实验中心,南宁530021

出　　处：《重庆医学》2014年第33期4450-4453,共4页Chongqing medicine

摘　　要：目的通过观察柔红霉素(DNR)作用后急性早幼粒细胞白血病(APL)细胞株HL-60敏感细胞中miRNA-21、miRNA-181d、miRNA-125b、let-7f和miRNA-27a5种miRNA的表达变化,探讨他们与HL-60细胞耐药性形成的相关性及其潜在的临床意义。方法常规培养HL-60和HL-60/A细胞。设置实验组、对照组1和对照组2,实验组使用小剂量DNR持续作用于HL-60敏感细胞。对照组1和2分别是HL-60/A DNR耐药细胞和不接受DNR持续作用的HL-60敏感细胞。采用四甲基偶氮唑盐比色(MTT)法检测DNR对HL-60细胞增的影响。采用实时荧光定量PCR(RT-PCR)检测miRNA-21等5种miRNA的表达量。结果 MTT实验结果:随着小剂量DNR持续作用时间的延长,DNR对HL-60敏感细胞的抑制作用减弱,并于加药后第10天左右,抑制率逐渐进入平台期。RT-PCR检测结果:与对照组2比较,对照组1耐药细胞中的miRNA-181d、miRNA-27a和let-7f的表达下调(P<0.05),而实验组敏感细胞中这些miRNA的表达于加药后的第7天开始下调(P<0.05)。相反,对照组1中的miRNA-21和miRNA-125b表达上调(P<0.05),而实验组敏感细胞中这2种miRNA的表达则于加药后的第7天开始上调(P<0.05)。结论 miRNA-21、miRNA-125b、miRNA-181d、miRNA-27a和let-7f5种miRNA表达的改变可能与HL-60细胞耐药性有关。Objective The purpose was to explore the relativity between the drug resistance of acute promyelocytic leukemia （APL） cell line HL‐60 and the expressions of miRNA‐21 ,miRNA‐181d ,miRNA‐125b ,let‐7f and miRNA‐27a in HL‐60 cells by an‐alyzing these 5 miRNA expressions in HL‐60 cells treated with daunorubicin （DNR） .The potential clinical significance of these 5 miRNAs expressions was also discussed .Methods The cell line HL‐60 and the doxorubicin resistant cell line HL‐60/A were rou‐tinely cultured .3 groups were set ,namely ,the experimental group and the controls group 1 and 2 .The experimental group was the HL‐60 cells were treated with DNR .The controls group 1 and 2 were the doxorubicin resistant HL‐60/A cells and the HL‐60 cells , respectively .MTT assay was carried out to examine the inhibition rate of HL‐60 cell treated with DNR .The expressions of miR‐NAs were analyzed by real time RCR（RT‐PCR） .Results MTT assay ：the results showed that with the extension of DNR treat‐ment ,DNR inhibition on HL‐60 cells became weak .DNR inhibition rate on cells graduated into a plateau on the 10th day after DNR treatment .RT‐PCR results ：comparing with the control group 2 ,the expressions of miRNA‐181d ,miRNA‐27a and let‐7f in the cells of the control 1 were obviously down‐regulated（P〈0 .05） .The expressions of these miRNAs in the sensitive cells of the experi‐mental group were down‐regulated on the 7th day after DNR treatment（P〈0 .05） .In contrast ,the expressions of miRNA‐21 and miRNA‐125b were up‐regulated in the cells of the control group 1（P〈0 .05） .The expressions of these miRNAs in the sensitive cells of the experimental group were up‐regulated on the 7th day after DNR treatment（P〈0 .05） .Conclusion These results indica‐ted that the change of the expression of miRNA‐21 ,miRNA‐125b ,miRNA‐181d ,miRNA‐27a and let‐7f might be related to drug re‐sistance of HL‐60 cells .

关 键 词：白血病 髓样 急性 微RNA 柔红霉素 白血病细胞株 耐药 

分 类 号：R733.7[医药卫生—肿瘤]