【摘要】 目的:研究沙利度胺对人红系细胞γ珠蛋白基因表达的作用及相关调控机制。方法:采用一步液体培养法培养人红系细胞,以不同浓度沙利度胺(50μmol/L、100μmol/L、200μmol/L、300μmol/L)、100μmol/L羟基脲(阳性对照)分别作用于人红系细胞24 h、48 h、72 h、96 h,并设无任何干预的人红系细胞为空白对照组。采用实时荧光定量PCR(qPCR)法检测γ珠蛋白基因及转录因子BCL11A、KLF1、KLF2、GATA1基因的表达水平,Western blotting法检测γ珠蛋白的表达量。结果:作用于人红系细胞72 h后,100μmol/L、200μmol/L、300μmol/L沙利度胺组γ珠蛋白基因相对表达量高于空白对照组,200μmol/L、300μmol/L沙利度胺组及阳性对照组γ珠蛋白表达量明显高于空白对照组(均P<0.05);与空白对照组比较,200μmol/L沙利度胺组BCL11A、KLF1基因表达明显下降,阳性对照组BCL11A基因表达下降,KLF2基因表达升高(P<0.05)。结论:沙利度胺能诱导体外人红系细胞γ珠蛋白基因表达,增加γ珠蛋白的生成,BCL11A、KLF1的下调可能是沙利度胺诱导γ珠蛋白基因表达的作用机制之一。更多还原

【Abstract】 Objective:To investigate the effect and mechanisms of thalidomide on the expression of γ globin gene in human erythroid cells.Methods:The human erythroid cells were treated with different concentrations(50 μmol/L,100 μmol/L,200 μmol/L and 300 μmol/L) of thalidomide or 100 μmol/L hydroxyurea(positive control) for 24 h,48 h,72 h and 96 h.Non-treatment cells were served as a blank control group.The mRNA expressions of γ globin,BCL11 A,KLF1,KLF2 and GATA1 were determined by fluorescence quantitative PCR(qPCR).The γ globin protein expression was detected by western blotting.Results:After 72 h of treatment,the mRNA expressions of γ globin in 100 μmol/L,200 μmol/L and 300 μmol/L thalidomide groups was significantly higher than that in the blank control group(P<0.05).The protein expression of γ globin was higher in 200 μmol/L and 300 μmol/L thalidomide groups and positive control group than that in the blank control group(P<0.05).Compared with the blank control group,the mRNA expressions of transcription factor BCL11 A and KLF1 were decreased in 200 μmol/L thalidomide group,the BCL11 A mRNA level was decreased while the KLF2 mRNA level was increased in the positive control group(P<0.05).Conclusion:Thalidomide could induce γ globin gene expression in erythroid cells and increase the production of γ globin.The mechanisms might be related to the down-regulation of BCL11 A and KLF1.更多还原