铁剥夺对化疗药物诱导HL-60细胞凋亡的影响

目的 :探讨铁剥夺诱导HL 60细胞及对化疗药物诱导HL 60细胞凋亡的影响。方法 :HL 60细胞与不同浓度的铁螯合剂 去铁胺 (DFO)、DFO加化疗药物、化疗药物、DFO加化疗药物及三氯化铁、三氯化铁共同培养 6h、12h、2 4h、48h。通过测定细胞活力 ,观察细胞形态学变化 ,应用流式细胞仪检测和DNA凝胶电泳等方法观察细胞凋亡 ;通过亲和免疫组化方法检测c myc基因表达 ,从而确定DFO及DFO与化疗药物联合应用对HL 60细胞的作用。结果 :DFO单用可降低HL 60细胞活力 ,抑制HL 60细胞增殖 ,诱导HL 60凋亡 ,并可使c myc基因表达增加 ,其作用强度随培养时间延长及DFO浓度增加而增加 ;DFO与化疗药物联合时 ,可增加化疗药物HL 60细胞凋亡的作用 ,该作用可被等摩尔的三氯化铁抵消。结论 :铁剥夺可影响HL 60细胞DNA的合成 ,诱导其凋亡 ,并提高HL 60细胞对化疗药物的敏感性。因此 ,铁剥夺可作为临床上白血病治疗或辅助治疗的方法之一 ,不适当补铁或升高血红蛋白将影响化疗疗效

Iron deprivation and apoptosis in HL-60 cells

Objective: This study aimed at investigation the effect of iron deprivation on HL 60 cells apoptosis.Methods: HL 60 cells were cultured with different concentrations of ferric (Ⅲ)chloride (FeCI 3),iron chelate desferrioxamine(DFO),cytosine arabinoside (Ara C), etoposide (VP 16), DFO+ Ara C/ VP 16, DFO+ FeCI 3+ Ara C/ VP 16 for 6 h,12 h,24 h,48 h.The proliferation of HL 60 cells was observed with cell viability assay. Apoptosis was determined with cell morphology ,flow cytometry ,DNA gel electrophoresis and the expression of c myc.Results: When HL 60 cells were incubated with different concentrations of FeCI 3,apoptosis percentages (APO%) was lower than those in control group.The lowest percentage was found at the concentration of 100 μmol/L FeCI 3.( P <0.05).In DFO group , APO% was much higher than those of control group ( P <0.05) and they were dose time dependant.The peak value appeared at 24 h at the concentration of100 μmol/L 150 μmol/L DFO.FeCI 3 +DFO+ chemotherapeutic drugs group with chemotherapeutic group there was no significant difference ( P >0.05).Expression percentage of c myc in DFO group was higher than that of control group At the concentration of 100 μmol/L DFO,( P <0.05).Conclusion: These results showed that iron could promote HL 60 cells growth and inhibit their apoptosis.Otherwise iron deprivation could induce HL 60 cell apoptosis.DFO disturbed the iron metabolism and inhibited DNA synthesis of HL 60.This action of DFO might enhance the susceptibility of HL 60 cell to apoptosis induced by chemotherapeutic drugs. Equal molar concentration of FeCI 3 could prevent the effect of DFO.Our report showed that the iron deprivation could have a place in the treatment of leukemia in combination with other anticancer agents .Therefore we suggest that as long as the hemoglobin level is maintained sufficiently for survival of leukemia patients ,red cell transfusion may not be necessary.