沙利度胺对重型β地中海贫血患者红系细胞γ珠蛋白基因表达及分化的作用研究  

Effect of thalidomide onγglobin gene expression anderythroid differentiation in erythroid cells of patients with severeβ-thalassemia

作　　者：黄婉雪 杨阳[1] 汪晓辉 赖永榕[1] HuangWanxue;Yang Yang;Wang Xiaohui;Lai Yongrong(Department of Hematology,The First Affiliated Hospital of Guangxi Medical University,Nanning530000,China)

机构地区：[1]广西医科大学第一附属医院,南宁530021

出　　处：《广西医科大学学报》2021年第1期9-15,共7页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81560024)

摘　　要：目的:探讨沙利度胺对重型β地中海贫血患者体外红系细胞γ珠蛋白基因(HBG)表达及分化的作用及相关调控机制。方法:以沙利度胺为实验组、羟基脲为阳性对照组、等体积溶剂二甲基亚砜(DMSO)为阴性对照组,分别作用于体外培养的重型β地中海贫血患者红系细胞。采用实时荧光定量PCR(qPCR)法检测HBG及参与调控HBG的转录因子的表达水平;蛋白免疫印迹法(Western blotting)检测γ珠蛋白的表达量;流式细胞仪检测各组细胞在分化不同阶段的成熟情况。结果:与DMSO组比较,沙利度胺组γ珠蛋白转录和翻译水平在分化第7天时逐渐增加(P<0.05);在分化末期,沙利度胺组细胞表面分化标记物表达下降(P<0.05);在分化过程中,调控珠蛋白转换的转录因子BCL11A在转录和翻译水平都降低(P<0.05);KLF1、GATA1在分化第7、第21天转录水平下降(P<0.05)。SOX6与TAL1在分化末期均下降(P<0.05)。结论:沙利度胺能提升体外重型β地中海贫血患者红系细胞γ珠蛋白转录及翻译水平,BCL11A、KLF1、GATA1等转录因子可能参与沙利度胺调控γ珠蛋白表达的过程,同时在分化末期,沙利度胺可能通过下调SOX6与TAL1减缓红系的分化,增加未成熟红细胞的增殖,调节血红蛋白转录,从而有效诱导γ珠蛋白。Objective:To investigate the effect and mechanisms of thalidomide on the expression ofγ-globin gene(HBG)and erythroiddifferentiation in erythroid cells ofpatients with severeβ-thalassemia.The erythroid cells of severeβ-thalassemia patientswere treated with thalidomide(experimental group),hydroxylurea(positive control group)and equal volume of dimethyl sulfoxide(DMSO,negative control group),respectively.The expression levels ofγ-globin gene and transcription factors involved in regulating HBG were detected by real-time fluorescence quantitative PCR(qPCR).The expression ofγ-globin was detected by Western blotting.Flow cytometry was used to detect the maturation of erythroid cells in each group at different differentiation phase.Results:Compared with the negative control group,the transcription and translation ofγ-globin in thalidomide group increased gradually on the 7th day of differentiation(P<0.05).At the end of differentiation phase,the expression of surface differentiation markers in thalidomide group decreased(P<0.05).The expression in the levels of transcription and translation of transcription factor BCL11A,which regulates globin conversion,was decreased(P<0.05).Transcription of KLF1 and GATA1 decreased on day 7 and 21 of differentiation(P<0.05).Both SOX6 and TAL1 decreased at the end of differentiation phase(P<0.05).Conclusion:Thalidomide can improve the transcription and translation of globin in erythrocytes ofβ-thalassemia major patients in vitro.BCLl1A,KLF1,GATA1 and other transcription factors may be involved in thalidomide regulation ofγ-globin expression.Moreover,at the late phase of differentiation,thalidomide may reduce the differentiation progression of erythroid cells differentiation by down-regulating SOX6 and TAL1,increase the proliferation of immature red blood cells,regulate hemoglobin transcription,and thus effectively induceγ-globin.

关 键 词：沙利度胺 Β地中海贫血 Γ珠蛋白 红系分化 

分 类 号：R556.61[医药卫生—血液循环系统疾病]