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目的探讨HL-60细胞铁池改变对凋亡相关基因表达的影响及可能的分子机制。方法实验分组为去铁胺(DFO)组、DFO+三氯化铁组及空白对照组,分别采用钙黄绿素检测HL-60细胞LIP、流式细胞术观察HL-60细胞凋亡和RT-PCR测定HL-60细胞bax、c-myc、rbmRNA表达。结果(1)不同浓度的DFO作用于HL-60细胞后,随培养时间延长及DFO浓度的增加,动态铁池降低,细胞生存率逐渐下降,显示一定的时间剂量依赖性。(2)不同浓度的DFO作用于HL-60不同时间后,细胞凋亡增加,高于对照组(P<0.05)。(3)RT-PCR结果显示,DFO能明显上调c-myc、rb和baxmRNA表达(P<0.05)。结论DFO通过螯合细胞内铁,降低HL-60细胞LIP,诱导细胞凋亡;诱导HL-60细胞凋亡的作用可能与其降低细胞动态铁池,上调细胞凋亡相关基因c-myc、rb、baxmRNA表达密切相关。
Objective To investigate the effect of alteration of iron pool in HL-60 cells on the expression of apoptosis-related genes and its possible molecular mechanism. Methods The experimental groups were divided into DFO group, DFO + ferric chloride group and blank control group. The LIP of HL-60 cells was detected by calcein and the apoptosis of HL-60 cells was observed by flow cytometry and RT-PCR HL-60 cells bax, c-myc, rbmRNA expression. Results (1) After different concentrations of DFO were added to HL-60 cells, the dynamic iron pool decreased and the cell survival rate decreased gradually with the prolongation of culture time and the increase of DFO concentration, indicating a certain time-dependent dose-dependent manner. (2) After treated with different concentrations of DFO for a long time, HL-60 cells apoptosis increased, which was higher than that of the control group (P <0.05). (3) The result of RT-PCR showed that DFO could obviously upregulate the expression of c-myc, rb and bax mRNA (P <0.05). CONCLUSION DFO can induce the apoptosis of HL-60 cells by chelating intracellular iron and reducing the LIP of HL-60 cells. The effect of DFO on HL-60 cells apoptosis may be related to the decrease of dynamic iron pool, upregulation of the expression of c-myc and rb, Bax mRNA expression is closely related.