microRNA-520a靶向MCM3抑制急性淋巴细胞白血病细胞增殖的研究  

Study on miR-520a targeting MCM3 inhibition on cell proliferation of acute B lymphoblastic leukemia cells

作　　者：张欢[1] 张国君[1] ZHANG Huan;ZHANG Guojun(First Department of Hematology,Affiliated Shengjing Hospital of China Medical University,Shenyang,Liaoning 110022,China)

机构地区：[1]中国医科大学附属盛京医院第一血液内科,沈阳110022

出　　处：《重庆医学》2021年第14期2366-2370,共5页Chongqing medicine

基　　金：辽宁省重点研发计划联合计划项目(2020JH2/10300134);沈阳市科学计划项目(18-014-4-20)。

摘　　要：目的阐述在急性B淋巴细胞白血病(B-ALL)细胞中microRNA(miR)-520a的表达和临床意义。方法在非恶性血液肿瘤患者和B-ALL患者骨髓淋巴细胞及健康人外周血淋巴细胞、B-ALL患者细胞中利用实时定量PCR检测miR-520a的表达情况;在人急性B淋巴细胞白血病细胞株(BALL-1)中转染miR-520a模拟物,CCK-8观察细胞增殖能力变化,实时定量PCR检测miR-520a的表达变化。生物信息学分析MCM3是miR-520a的靶基因并预测位点;使用双萤光素酶报告基因检测miR-520a对MCM3的转录活性影响;在miR-520a mimics转染组中使用MCM3过表达质粒进行回补实验,观察MCM3蛋白和细胞增殖变化。结果同正常组织/细胞相比,B-ALL患者细胞/BALL-1细胞系中miR-520a表达水平显著降低(P<0.05),miR-520a过表达能够使BALL-1细胞增殖能力降低(P<0.05);miR-520a过表达降低MCM3的蛋白表达水平(P<0.05);通过双萤光素酶实验验证miR-520a能够作用MCM3的3′UTR区;miR-520a mimics能够使BALL-1细胞增殖能力下降,同时过表达MCM3能够提高细胞增殖能力。结论miR-520a可能通过下调MCM3的表达抑制B-ALL细胞增殖能力。Objective To elaborate the expression and clinical significance of miR-520a in acute B-lymphoblastic leukemia(B-ALL)cells.Methods The expression level of miR-520a in the bone marrow lymphocytes of the patients with non-malignant blood tumor and the patients with B-ALL,peripheral blood lymphocytes of normal people and cells of the patients with B-ALL was detected by real-time fluorescent quantitative PCR.The miR-520a mimics were transfected into BALL-1 cells.CCK-8 was used to observe the cell proliferation ability change and the expression of miR-520a was detected by real-time quantitative PCR.The bioinformatics method was used to analyze MCM3 as the target gene of miR-520a and predict its action site;the double luciferase reporter gene system was used to detect the effect of miR-520a on MCM3 transcriptional activity;in the miR-520a mimics transfection group,the MCM3 overexpression plasmid was used to conduct the complement experiment for observing the change of MCM3 protein and cell proliferation.Results Compared with the normal tissues/cells,the expression level of miR-520a in bone marrow lymphocytes/BALL-1 cell lines of the patients with B-ALL was significantly decreased(P<0.05),and the overexpression of miR-520a could reduce the proliferation ability of BALL-1 cells(P<0.05),and the protein expression level of MCM3 was decreased by miR-520a overexpression(P<0.05).The double luciferase assay verified that miR-520a could act the 3′UTR region of MCM3.miR-520a mimics could decrease the proliferation ability of BALL-1 cells,meanwhile the overexpression of MCM3 could increase the cell proliferation ability.Conclusion miR-520a may inhibit the proliferation of B-ALL cells by down-regulating the expression of MCM3.

关 键 词：前体细胞淋巴母细胞白血病淋巴瘤 细胞增殖 miR-520a MCM3 

分 类 号：R552[医药卫生—血液循环系统疾病]