5-氮杂-2’-脱氧胞苷和曲古抑菌素A对1,2,4-苯三醇调控K562细胞GATA结合蛋白1表达水平的影响  

Effect of 5-aza-2'-deoxycytidine and trichostatin A on the expression of GATA-1 in K562 cells induced by 1,2,4-benzenetriol

作　　者：崔宁轩 赵霄[1] 王颖[1] 余春红[1] 易宗春[1] CUI Ning-xuan;ZHAO Xiao;WANG Ying;YU Chun-hong;YI Zong-chun(School of Biological Science and Medical Engineering, Beihang University, Beijing 100083, China)

机构地区：[1]北京航空航天大学生物与医学工程学院,北京100083

出　　处：《毒理学杂志》2018年第2期93-97,共5页Journal of Toxicology

基　　金：国家自然科学基金(81573192和81072325)

摘　　要：目的 GATA-1是在红系分化过程中发挥关键作用的转录因子,实验观察1,2,4-苯三醇对K562细胞GATA-1基因mRNA表达的影响,以及DNA甲基化抑制剂5-氮杂-2’-脱氧胞苷(5-aza-CdR)和组蛋白去乙酰化酶抑制剂曲古抑菌素A(TSA)的干预作用。方法培养的K562细胞先经5、10和20μmol/L的1,2,4-苯三醇处理72 h,然后经氯化高铁血红素诱导后,应用联苯胺染色法检测细胞血红蛋白合成情况,采用反转录PCR检测细胞GATA-1的mRNA表达水平。培养的K562细胞先经20μmol/L的1,2,4-苯三醇处理24或48 h后,或加入5-aza-CdR共同孵育48 h,或加入TSA共同孵育24 h,然后经氯化高铁血红素诱导后,采用反转录PCR检测细胞GATA-1的mRNA表达水平。结果当K562细胞经5、10和20μmol/L的苯三醇处理后,氯化高铁血红素诱导下的GATA-1mRNA表达水平分别只有对照组的91.1%、52.7%和23.2%,红系分化率分别只有对照组的93.3%、85.9%和65.5%。如果在20μmol/L的苯三醇处理24 h后加入5-Aza-CdR共同孵育48 h,氯化高铁血红素诱导的GATA-1mRNA表达水平相对于单纯苯三醇处理组有明显回升,达到对照组水平的80.0%。如果在20μmol/L的苯三醇处理48 h后加入TSA共同孵育24 h,氯化高铁血红素诱导的GATA-1mRNA表达水平相对于单纯苯三醇处理组有明显回升,达到对照组水平的78.0%。结论 1,2,4-苯三醇可引起K562细胞GATA-1基因转录抑制,而DNA甲基化和组蛋白去乙酰化的表观遗传修饰方式的改变可能参与其中。Objective GATA-1 is a transcription factor that plays a key role in erythroid differentiation. The purpose of this experiment is to investigate the effect of 1,2,4-benzenetriol on GATA-1 expression in K562 cells,and the impacts of DNA methylation inhibitor 5-aza-2＇-deoxycytidine(5-aza-CdR） and histone deacetylation inhibitor trichostatin A(TSA） in this process. Methods In this study,K562 cells were treated with 1,2,4-benzenetriol which concentration was 5 or 10 or 20 μmol/L for 72 h. After K562 cells treated with1,2,4-benzenetriol,the hemin-induced hemoglobin synthesis was detected by benzidine staining,and reverse transcription PCR was used to detect the mRNA level of GATA-1. After K562 cells treated with 1,2,4-benzenetriol which concentration was 20 μmol/L for 24 h,5-aza-CdR was added to incubate for 48 h; After K562 cells treated with 1,2,4-benzenetriol which concentration was 20 μmol/L for 48 h,TSA was added to incubate for 24 h. After these co-incubation,K562 cells were induced by hemin and reverse transcription PCR was used to detect the mRNA level of GATA-1. Results After K562 cells exposure to 1,2,4-benzenetriol,the hemin-induced expression of GATA-1 genes was91. 1% of the control when the concentration of 1,2,4-benzenetriol was 5 μmol/L; the hemin-induced expressions of GATA-1 genes was52. 7% of the control when the concentration of 1,2,4-benzenetriol was 10 μmol/L; the hemin-induced expressions of GATA-1 genes was23. 2% of the control level when the concentration of 1,2,4-benzenetriol was 20 μmol/L; After K562 cells exposure to 1,2,4-benzenetriol which concentration was 20 μmol/L for 24 h,then 5-aza-CdR was added to incubate for 48 h,the hemin-induced expressions of GATA-1 genes were significantly increase,which was 80. 0% of the control. After K562 cells exposure to 1,2,4-benzenetriol which concentration was20 μmol/L for 48 h,then 5-aza-CdR was added to incubate for 24 h,the hemin-induced expressions of GATA-1 genes were significantly increase,which was 78. 0% of the control. Conclusion 1

关 键 词：1 2 4-苯三醇 红系分化 GATA-1 DNA甲基化 组蛋白去乙酰化 

分 类 号：R992[医药卫生—毒理学]