miR-181a通过调节Bcl-2逆转白血病细胞的耐药作用

The Effect of miR-181a on Drug Resistance of Leukemia Cells by Regulating Bcl-2 Expression

目的研究微小RNA-181a(miR-181a)和B淋巴细胞瘤-2基因(Bcl-2)在急性粒细胞白血病(AML)老年患者临床标本、人早幼粒急性白血病细胞株(HL60)及人早幼粒急性白血病细胞耐阿霉素株(HL60/ADR)中的表达,并探讨白血病化疗耐药与miR-181a之间的关系。方法收集2019年1月至2021年3月经泰州市人民医院血液内科确诊的22例AML初诊者、12例完全缓解者、11例复发者和10例正常者资料。利用脂质体2000(Lipofectamine2000)转染试剂以转染寡核苷酸探针,实时荧光定量PCR检测miR-181a和Bcl-2的表达水平,Western Blot方法检测Bcl-2的蛋白表达水平,MTT方法检测HL60和HL60/ADR细胞的存活率。结果与缓解者比较,AML初诊者和复发者样本中miR-181a表达显著下降,Bcl-2的表达显著升高,差异均有统计学意义(P<0.05);同样地,与HL60细胞比较,HL60/ADR细胞中miR-181a表达显著下降,Bcl-2的表达显著升高,差异均有统计学意义(P<0.01)。结论miR-181a可通过对Bcl-2表达的调节以参与白血病耐药的形成,miR-181a可能成为逆转白血病耐药的重要靶点。

Objective To investigate the expression of microRNA(miR-181a)and B-lymphocytoma-2 genes(Bcl-2)in clinical specimens of elderly patients with acute myoloid leukemia(AML),in human promyeloid acute leukemia cell line(HL60)and in human promyeloid acute leukemia cell line adriamycin resistant(HL60/ADR),and discuss the relationship between miR-181a and drug resistance in leukemia treatment.Methods Treatment-naive,CR or relapsed AML patients hospitalized in the Department of Hematology,Taizhou People’s Hospital from January 2019 to March 2021 were enrolled,along with healthy volunteers as controls.Oligonucleotide probes were transfected in HL60 and HL60/ADR cells using Lipofectamine 2000.Real-time fluorescent quantitative PCR was used to detect the expression of miR-181a and Bcl-2.Protein expression of Bcl-2was analyzed via Western blot.The viability of HL60 and HL60/ADR cells was evaluated by MTT assay.Results A total of 22 AML treatment-na?ve patients,12 CR patients,11 relapsed patients and 10 healthy volunteers were enrolled.Patients with complete recession had significantly lower expression of miR-181a and higher expression of Bcl-2 than those in treatment-naive and relapsed patients(P<0.05,respectively).Similarly,miR-181a expression was reduced and Bcl-2 expression was elevated in HL60/ADR cells,compared with HL60 cells(P<0.01,respectively).Upregulating the expression of miR-181a led to increase inhibition rate of adriamycin on HL60/ADR cells and inhibited Bcl-2 expression(P<0.01,respectively).Meanwhile,knock-down of miR-181a led to reduced inhibition rate of adriamycin on HL60 cells and elevated Bcl-2 expression(P<0.01,respectively).Conclusion MiR-181a was involved in the mechanism of drug resistance in leukemia by regulating Bcl-2 expression,which might provide a potential novel target for overcoming drug-resistance.