HL-60细胞铁池变化与凋亡相关基因关系的初步研究

Interacations between the genes of apoptosis and the labile iron pool in HL-60 cells

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 explore the effect of iron chelators on expression of apoptosis relating-genes in HL-60 leukemia cells. Methods We used exponentially growing HL-60 and K562 cells （ 1 × 10^6/ml） in experiment. The study groups were divided as following： DFO group, iron ＋ DFO and control group. Following indices were detected which included viability by typanblue, apoptosis by morphological study, flow eytometry （FCM） assay, expression of rb, c-myc, baxmRNAby RT-PCR, The intracellular LIP was measured with a fluorimetrie assay using the metalsensitive probe calcein-AM. Results the expression levels of Rb and c-myc were increased significantly by DFO （50 μmol/L and 100 μmol/L） （P 〈0. 05）, but the extent of the increase was predominant when DFO was 100 μmol/L（P 〈0. 01 ）. Conclusions The correlation was a negative between cellular LIP and expression of apoptosis-related increasing the expression of HL-60 cells may be one of mechanisms DFO induced apoptosis in leukemia cells.