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目的探讨不同铁状态下IRP2 mRNA和TfR mRNA在HL-60细胞中的表达以及二者之间的关系。方法将FeCl3或去铁胺(Desferrioxamine,DFO)加入含10%胎牛血清的RPMI-1640培养液中,使HL-60细胞处于缺铁或富铁的状态下进行培养。于细胞培养后第12、24和48小时收集各组细胞,提取总RNA。采用RT-PCR半定量法测定IRP2 mRNA和TfR mRNA的相对表达量。结果①各实验组之间IRP2 mRNA的表达量变化不大,无明显差异(F组间=1.199,P>0.05)。细胞培养时间对IRP2 mRNA的表达有影响,差异有显著性(F组内=43.418,P<0.05),表现为随时间的延长,IRP2 mRNA的表达降低。②各实验组之间TfR mRNA的差异有显著性(FW-S=7.184,F组间=113.926,P<0.01)。在加入DFO的两个实验组中TfR mRNA表达升高。在加铁的两个实验组中,与对照组相比较,在12h时TfR mRNA的表达量上升,24h时达到高峰,之后迅速下降。结论①铁或去铁胺对HL-60细胞IRP2 mRNA的表达无直接影响。②当细胞内铁降低时,TfR mRNA的基因表达则增加;当细胞内铁增加时,TfR mRNA的基因表达则降低。
Objective To investigate the expression of IRP2 mRNA and TfR mRNA in HL-60 cells under different iron status and their relationship. Methods FeCl3 or Desferrioxamine (DFO) was added into RPMI-1640 culture medium containing 10% fetal calf serum to culture HL-60 cells in the condition of iron deficiency or iron deficiency. At 12, 24 and 48 hours after cell culture, each group of cells were collected and total RNA was extracted. The relative expression levels of IRP2 mRNA and TfR mRNA were determined by semi-quantitative RT-PCR. Results ① There was no significant difference in the expression of IRP2 mRNA between experimental groups (F = 1.199, P> 0.05). The cell culture time had an impact on the expression of IRP2 mRNA, the difference was significant (F = 43.418, P <0.05), which showed that the expression of IRP2 mRNA decreased with the prolongation of time. ② The difference of TfR mRNA between experimental groups was significant (FW-S = 7.184, F = 113.926, P <0.01). TfR mRNA expression increased in both experimental groups with addition of DFO. Compared with the control group, the expression of TfR mRNA in the two experimental groups of iron increased at 12 hours, reached a peak at 24 hours and then decreased rapidly. Conclusion ① Iron or deferoxamine has no direct effect on the expression of IRP2 mRNA in HL-60 cells. ② When the intracellular iron decreased, TfR mRNA gene expression increased; when intracellular iron increased, TfR mRNA gene expression decreased.