RNA甲基化“阅读器”蛋白YTHDF2在急性髓系白血病中的作用  

Role of RNA methylation reader YTHDF2 in the development of human acute myeloid leukemia

作　　者：曹丽霞[1] 陈连香[1] CAO Li-xia;CHEN Lian-xiang(Dept.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,内蒙古呼和浩特010059

出　　处：《基础医学与临床》2018年第12期1727-1732,共6页Basic and Clinical Medicine

摘　　要：目的探讨RNA甲基化修饰(m6A)特异性"阅读器"蛋白YTHDF2在急性髓系白血病人外周血单个核细胞中的表达水平改变,并研究YTHDF2对急性髓系白血病细胞增殖、分化的影响。方法分别收集急性髓系白血病患者以及正常人外周血的单个核细胞,实时定量PCR检测YTHDF2 mRNA在其中的表达差异;分别使用特异性靶向YTHDF2的siRNA和对照siRNA转染急性髓系(单核系)白血病细胞系THP-1细胞;通过CCK-8法检测细胞增殖;通过流式细胞计量术检测细胞向单核系分化的情况;使用放线菌素D处理结合实时定量PCR检测YTHDF2对其下游靶基因MZF1 mRNA稳定性的影响;用Western blot检测下游靶基因MZF1蛋白表达。结果 YTHDF2 mRNA在急性髓系白血病患者中的表达水平显著高于对照组正常人;敲低内源性YTHDF2的表达可以抑制THP-1细胞的增殖(P<0. 05);同时促进PMA诱导的THP-1单核细胞分化; YTHDF2可通过影响靶基因MZF1 mRNA的稳定性,抑制MZF1蛋白水平的表达(P<0. 05)。结论 YTHDF2靶向抑制MZF1的表达,在急性髓系白血病发生发展中发挥促癌功能。Objective To study the expression and function of RNA methylation reader YTHDF2 in the development of human acute myeloid leukemia(AML).Methods The peripheral blood mononuclear cells(PBMCs)were isolated from AML patients and controls were used to detect the relative expression of YTHDF2 mRNA by quantitative RT-PCR analysis;THP-1 cells were transfected with siRNAs specific to YTHDF2 or control.The effects of YTHDF2 knock-down on THP-1 cell proliferation were examined by CCK-8(P<0.05).The effects of YTHDF2 knock-down on THP-1 cell monocytic differentiation were examined by FACS analysis.The influence of YTHDF2 on target gene MZF1 mRNA stability was determined by quantitative RT-PCR after AcD treatment.The influence of YTHDF2 on target gene MZF1 protein expression was determined by Western blot(P<0.05).Results The expression of YTHDF2 mRNA was up-regulated in the PBMCs from AML patients compared to the normal controls.Knock-down of YTHDF2 reduces THP-1 cell proliferation and promotes its monocytic differentiation induced by PMA.YTHDF2 could regulate MZF1 expression by affecting the stability of MZF1 mRNA.Conclusions YTHDF2 may act as oncogene in AML by controlling the expression of its target gene MZF1.

关 键 词：YTHN(6)-甲基腺苷核糖核酸结合蛋白2 RNA甲基化修饰 髓样锌指蛋白1 急性髓系白血病 

分 类 号：R365[医药卫生—病理学]