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作 者:陈连香[1] 曹丽霞[1] CHEN Lian-xiang;CAO Li-xia(Department of Hematology,the Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010059,China)
机构地区:[1]内蒙古医科大学附属医院血液内科
出 处:《基础医学与临床》2019年第9期1259-1264,共6页Basic and Clinical Medicine
基 金:内蒙古自治区自然科学基金[2017MS(LH)0835];内蒙古医科大学附属医院科研项目(YB004)
摘 要:目的探讨miR-150在人慢性髓系白血病细胞系K562中的功能和作用机制。方法以临床收集的慢性髓系白血病患者、和健康人外周血中的单个核细胞为实验材料;实时定量PCR检测miR-150和c-Myb mRNA的表达水平;使用miR-150模拟物转染K562细胞;通过CCK-8法检测K562细胞增殖;通过流式细胞计量术检测K562细胞周期;荧光素酶报告基因实验结合Western blot检测miR-150对其下游靶基因c-Myb的调控作用。结果慢性髓系白血病患者与健康人相比,miR-150表达降低,而c-Myb表达升高;过量表达miR-150可以抑制K562细胞的增殖(P<0.05),同时,阻滞K562细胞周期的运行;miR-150可以下调靶基因c-Myb的表达。结论miR-150在慢性髓系白血病中表达下调,其作用机制是通过抑制癌基因c-Myb的表达抑制白血病细胞的增殖。Objective To investigate the function and mechanism of miR-150 in human chronic myeloid leukemia cell line K562.Methods Clinic CML patients and normal controls were recruited.The peripheral blood mononuclear cells from above CML and controls were isolated.Quantitative PCR analysis was used to determine the expression level of miR-150 and c-Myb mRNA;miR-150 mimic and negative control were transfected into K562 cells,respectively.CCK-8 analysis was performed to examine K562 cell proliferation.FACS analysis was used to detect cell cycle.Dual-luciferase assay and Western blot were used to detect the influence of miR-150 on target gene c-Myb.Results When compared to the normal control,the miR-150 level was down-regulated in CML patients,whereas c-Myb expression was up-regulated in CML.Over-expression of miR-150 inhibited K562 cell proliferation and cell cycle progression.miR-150 may reduce c-Myb expression in K562 cells.Conclusions miR-150 is down-regulated in CML,and it can inhibite K562 cell growth by expression of oncogene c-Myb.
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