G-CSF增加三氧化二砷对骨髓增生异常综合征细胞系SKM-1的细胞毒作用

G-CSF enhances the cytotoxicity of arsenic trioxide against myelodysplastic syndromes cell line SKM-1

目的探讨粒细胞集落刺激因子(G-CSF)增强三氧化二砷(As2O3)诱导骨髓增生异常综合征(MDS)细胞株SKM-1细胞凋亡作用。方法将不同浓度G-CSF(10、20、40ng/mL)与SKM-1细胞共育48h,以不加G-CSF为空白对照组,碘化丙锭细胞核染色法结合流式细胞术分析细胞周期变化。继之以20ng/mL G-CSF预处理细胞24h,不加G-CSF组为对照组,然后将不同浓度的As2O3(2.5、5.0、10.0μmol/L)加入实验组和对照组共培养48h,分别采用Cell Counting Kit(CCK-8)检测细胞增殖和Annexin V/PI试剂盒结合流式细胞术检测细胞凋亡水平。结果 20ng/mL G-CSF为促进SKM-1细胞周期向S期转化的更好浓度(P<0.05);CCK-8检测显示As2O3+G-CSF组较As2O3单药组抑制细胞生长更明显(P<0.05);凋亡结果表明As2O3+G-CSF组细胞凋亡率高于As2O3组(P<0.05)。结论 G-CSF动员SKM-1细胞进入S期可能是其增强As2O3的诱导凋亡作用的机制之一。

Objective To investigate the effect of granulocyte colony-stimulating factor （G-CSF） on arsenic trioxide （As 2O 3） induced apoptosis in myelodysplastic syndrome （MDS） cell line SKM-1. Methods Different concentrations of G-CSF （10,20,40 ng/mL） were co-cultured with SKM-1 cells for 48 h,with no G-CSF as the blank control group,flow cytometry was used to determined the cell cycle combined with propidium iodide staining.The cells were pretreated with 20 ng/mL G-CSF for 24 h,and no G-CSF as the control group.Then different concentrations of As 2O 3 （2.5,5.0,10.0 μmol/L） were added to the experimental group and the control group for co-culture for 48 h,Cell Counting Kit （CCK-8） was used to detect cell proliferation and Annexin V/PI kit combined with flow cytometry were used to detect apoptosis. Results 20 ng/mL G-CSF was a better concentration to promote SKM-1 cell cycle to S phase （ P 〈0.05）;CCK-8 assay showed that As 2O 3＋G-CSF inhibited the cell growth more significantly than that of the As 2O 3 group （ P 〈0.05）;apoptosis results showed that the apoptosis rate of the As 2O 3＋G-CSF group was significantly higher than that of the As 2O 3 group （ P 〈 0.05 ）. Conclusion G-CSF can enhance As 2O 3-induced apoptosis by mobilizing SKM-1 cells into S phase.