氯化高铁血红素诱导K562细胞中β-珠蛋白基因表达的研究 A Study of Inductive Effect of Hemin on Expression of theβ-globin Genes in K562 Cells

以绿色荧光蛋白（EGFP）基因为报道基因，构建了在β-珠蛋白启动子驱动及HS2元件调控下的重组表达载体HG，用脂质体转染法将其转染到K562细胞中，并用RT-PCR方法及流式细胞仪检测氯化高铁血红素(Hm)对K562细胞中β-珠蛋白基因表达及重组载体HG在K562细胞中瞬时表达的影响。结果显示：用30μmol/L Hm诱导K562细胞24、48及72h后，不仅其γ-珠蛋白mRNA水平升高，其β-珠蛋白mRNA水平也明显上升，而且这种诱导作用在诱导24、48h后比较明显；Hm还可增强重组表达载体HG在K562细胞中的瞬时表达。提示Hm诱导红系分化的机理可能与γ→β-珠蛋白基因的转换机制相关。 Abstract: The recombinant plasmid HG was constructed,in which the reporter gene encoding the enhanced green fluorescent protein (EGFP) was driven by the β-globin promoter and regulated under the HS2 element.The inductive effect of hemin on the expression of the β-globin gene and transiently transfected β-globin genes in K562 cells was analysed by FACS as well as RT-PCR method.The results showed that the level of γ and β-globin gene mRNA in K562 cells increased significantly after 24,48 and 72 hours induced with 30 μmol/LHm.And this inductive effect was sronger after 24 and 48 hours.Furthermore,the transient expression of plasmid HG in K562 cells increased significantly with hemin induction.Th