【摘要】 目的　探讨铁剥夺对HL 6 0细胞凋亡及对化疗药物诱导HL 6 0细胞凋亡的影响。方法　HL 6 0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 6 0细胞的作用。结果　DFO单用可降低HL 6 0细胞活力、抑制HL 6 0细胞增殖、诱导HL 6 0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 ,可增加化疗药物诱导HL 6 0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 6 0细胞DNA的合成 ,诱导其凋亡 ,并提高HL 6 0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。更多还原

【Abstract】 Objective Iron is an essential element for cell growing including tumor cells. This study aimed at investigating the effect of iron deprivation on HL-60 cell apoptosis. Methods HL-60 cells were cultured with different concentrations of ferric (Ⅲ) chloride (FeCL 3), iron chelate-desferrioxamine (DFO), cytosine arabinoside (Ara-C), etoposide (VP-16), DFO+Ara-C/VP-16, DFO+FeCL 3+Ara-C/VP-16 for 6 hr, 12 hr, 24 hr and 48 hr. 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 FeCL 3, apoptosis percentages (APO%) was lower than those in control group. The lowest percentage was found at the concentration of 100 μmol/L FeCL 3. APO% of HL-60 in this group at 12 hr, 24 hr and 48 hr were 2.2±0.6, 3.7±1.6 and 6.1±1.3 respectively, while APO% in control group were 4.6±0.8, 5.2±1.4 and 10.3±1.9 (P<0.05). In DFO group, APO% was much higher than those of control group (P<0.05) and they were dose-time dependent. The peak value appeared at 24 hr at the concentration of 100 μmol/L- 150 μmol/L DFO. At the concentration of 100 μmol/L DFO, APO% at 6 hr, 12 hr, 24 hr and 48 hr were 12.5±2.6, 21.4±5.9, 41.4±6.3 and 25.6±12.4 respectively (P<0.05). After co-culture with Ara-C and DFO for 6 hr, 12 hr and 24 hr, APO% were 18.0±5.7, 38.3±5.0 and 43.5±8.4 respectively, while APO% in Ara-C only group were 8.6±2.7, 26.0±4.3 and 30.7±5.3 (P<0.05). In comparison of FeCL 3+DFO+chemotherapautic 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, expression percentage at 6 hr, 12 hr and 24 hr were 4.1±1.2, 18.3±4.1 and 53.4±7.0 respectively (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 cells. This action of DFO might enhance the suseeptbility of HL-60 cells to apoptosis induced by chemotherapeutic drugs. Equal molar concentration of FeCL 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.更多还原