摘要
为了研究细胞可变铁池(labileironpool,LIP)的检测方法,用不同浓度的去铁胺(DFO)与K562及HL-60细胞共同培养0、6、12、24、48小时,用钙荧光素(caicein)负荷K562及HL-60细胞,荧光分辨仪检测细胞荧光量;用快速铁螯合剂螯合铁,使恢复荧光,然后计算2次荧光差值。结果表明:浓度为50和100μmol/L的DFO作用于K562及HL-60细胞12小时后,荧光值明显高于对照组(P<0.05),且LIP的荧光差值(ΔLIP)随DFO作用时间的延长及浓度增加而降低,即呈一定的时间及剂量依赖性。结论:DFO通过螯合细胞内铁降低了白血病细胞LIP;calceinAM荧光可作为检测细胞LIP变化的比较可靠的方法。
To explore a rapid and easy method to detect labile iron of pool （LIP） in cells, HL-60 and K562 cells were cultured at a concentration 1×10^6/ml in RPMI 1640 containing 10% heat- inactivated fetal bovine serum. The iron deprivation was induced by adding de sferrioxamine （DFO） 10 100μmol/L for 0-48 hours. The intracellular LIP was measured by probe calcein-AM. Calcein fluorescence was monitored in 1420 multilabel counter. The results indicated that when HL-60 and K562 cells were incubated with different concentrations of DFO, the calcein fluorescence intensity was higher than that of control group at 12, 24 and 48 hours （ P〈0.05 ）. Fluorescence value of representing LIP in DFO groups was lower than that in the control group. In conclusion, DFO can decrease LIP in leukemia cells. The approach used in this study may provide a simple and reliable method for detection of intracellular iron homeostasis.
出处
《中国实验血液学杂志》
CAS
CSCD
2006年第3期468-470,共3页
Journal of Experimental Hematology
基金
国家自然基金资助项目(编号:30370601)
