重组融合蛋白IL-3-PE38KDEL对急性髓性白血病细胞增殖和凋亡的影响

目的观察重组融合蛋白IL-3-PE38KDEL对急性髓性白血病(Acute myelocytic leukemia,AML)细胞HL-60(IL-3R阳性)和NB4(IL-3R阴性)增殖和凋亡的影响。方法用不同浓度的IL-3-PE38KDEL分别作用HL-60和NB4细胞,MTT法检测细胞的增殖抑制作用,计算抑制率及半数抑制浓度(IC50)。将HL-60和NB4细胞分别分为IL-3-PE38KDEL组和IL-3+IL-3-PE38KDEL组,IL-3+IL-3-PE38KDEL组加入IL-3(100 ng/ml),孵育4 h后,IL-3-PE38KDEL组和IL-3+IL-3-PE38KDEL组均加入IC50浓度的IL-3-PE38KDEL,计算细胞增殖抑制率;流式细胞术检测2种AML细胞细胞周期的变化及凋亡情况。结果 IL-3-PE38KDEL对HL-60和NB4细胞的增殖均有抑制作用,且呈浓度依赖性,对HL-60细胞的抑制作用强于NB4细胞(P<0.05),IC50值分别为2.5μg/ml和259.2μg/ml。当IL-3-PE38KDEL浓度为IC50时,经IL-3处理后,对HL-60和NB4细胞增殖的抑制率均值分别为32.20%和49.00%。经IL-3-PE38KDEL作用后的HL-60细胞大部分被阻滞于G1/S期。HL-60细胞IL-3-PE38KDEL组的细胞凋亡率均值(19.71%)显著高于IL-3+IL-3-PE38KDEL组(9.39%)(P<0.05),NB4细胞两组差异无统计学意义(P﹥0.05)。结论 IL-3-PE38KDEL能抑制IL-3R阳性的AML细胞HL-60增殖,并诱导其凋亡,但对IL-3R阴性的AML细胞NB4的抑制作用较弱。

Effect of Recombinant Fusion Protein IL-3-PE38KDEL on Proliferation and Apoptosis of Acute Myelocytic Leukemia Cells

Objective To observe the effect of recombinant fusion protein IL-3-PE38KDEL on the proliferation and apoptosis of acute myelocytic leukemia(AML) HL-60(IR-3R-positive) and NB4(IL-3R-negative) cells.Methods HL-60 and NB4 cells were treated with various concentrations of IL-3-PE38KDEL separately,and determined for proliferation levels by MTT method,based on which the inhibiting rate and median inhibiting concentration(IC50) to proliferation were calculated.HL-60 and NB4 cells were divided into IL-3-PE38KDEL and IL-3 + IL-3-PE38KDEL groups separately.The cells IL-3-PE38KDEL were added with IL-3-PE38KDEL at the IC50,while those in IL-3 + IL-3-PE38KDEL group were added with 100 ng / ml IL-3 and incubated for 4 h before addition of IL-3-PE38KDEL.The inhibiting rates of cell proliferation in various groups were calculated,and the cell cycles and apoptosis were determined by flow cytometry.Results IL-3-PE38KDEL showed dose-dependent inhibitory effect on both the proliferations of HL-60 and NB4 cells.However,the inhibiting rate of proliferation of HL-60 cells was significantly higher than that of NB4 cells(P < 0.05),while the IC50 were 2.5 and 259.2 μg/ml respectively.The mean inhibiting rates of IL-3-PE38KDEL at the IC50 to the proliferations of HL-60 and NB4 cells after treatment with IL-3 were 32.20% and 49.00% respectively.A majority of HL-60 cells after treatment with IL-3-PE38KDEL were arrested at G1 / S phase.The mean apoptosis rate of HL-60 cells in IL-3-PE38KDEL group(19.71%) was significantly higher that in IL-3 + IL-3-PE38KDEL group(P < 0.05),while those of NB4 cells in the two groups showed no significant difference(P > 0.05).Conclusion IL-3-PE38KDEL inhibited the proliferation and induced the apoptosis of IL-3R-positive AML HL-60 cells,while showed weak inhibitory effect on IL-3R-negative AML NB4 cells.