低氧对红系转录因子表达的影响

Effect of hypoxia on the expression of erythroid transcription factors

目的通过观察慢性高原病患者骨髓红系统K562细胞中HIF-1α及红系转录因子的表达水平,探讨低氧和红系转录因子对红系造血的影响。方法利用磁珠免疫分选技术获得慢性高原病骨髓单个核细胞中CD235a细胞作为观察组,同时选择白细胞轻度减少患者作为对照组,通过RT-PCR法检测骨髓CD235a细胞中HIF-1α及红系转录因子mRNA的表达,将过表达及干扰HIF-1α的慢病毒转染后的K562细胞在常氧状态下培养72 h后收集细胞,分别作为过表达组和干扰组,用RT-PCR检测红系转录因子的表达水平。结果结果显示观察组HIF-1α和GATA1 mRNA表达高于对照组,而NFE2、c-MYB、FOG-1、LMO2、β-globin和α-globin的mRNA水平无显著差异。慢病毒转染K562细胞最佳转染复数MOI为10,转染效率为50%左右,经过嘌呤霉筛选后转染阳性细胞达90%以上。过表达组中GATA1、NFE2、FOG-1和β-globin mRNA表达高于干扰组。上述差异具有统计学意义(P<0.05)。结论低氧-HIF-1-红系特异基因(GATA-1、FOG-1和β-globin)表达轴在低氧诱导红系造血过程中发挥着重要的调控作用。

Objective The purpose of this study was to observe the expression of HIF-1α and erythroid transcription factors in bone marrow erythrocytes in chronic mountain sickness and K562 cells,and to investigate the effects of hypoxia and erythroid transcription factors on the erythrocytes hematopoiesis.Methods The C.D235 a cells in bone marrow mononuclear cells of chronic mountain sickness were obtained by using magnetic activated cell sorting as CMS group and healthy persons as control group.The expression HIF-1α and erythroid transcription factor mRNA were tested by RT-PCR in bone marrow CD235 a cells of CMS.The K562 cells were transfected by interfered with HIF-1α gene.Transfected K562 cells with carried and interfered with HIF-1α gene were used as over-expression and interference group,respectively.The cells were harvested after cultured under normoxic conditions for 72 hours.The expression of HIF-1α and erythroid transcription factors were detected by RT-PCR.Results The results showed that the expression of HIF1-and GATA1 mRNA in the CMS group were higher than that in the control group,while there was no significant difference in the mRNA levels of NFE2,c-MYB,FOG-1,LMO2,β-globin,and α-globin.The expression of GATA1,NFE2,FOG-1 and β-globin mRNA in the over-expression group was significantly higher than the interference group.The above differences were statistically significant(P <0.05).Conclusion The axis of hypoxia-HIF-1-erythroid-specific genes(GATA-1,FOG-1 and β-globin)played an important regulatory role in hypoxia-induced erythroid hematopoiesis.