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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。
Objective To observe the effect of recombinant fusion protein IL-3-PE38KDEL on the proliferation and apoptosis of HL-60 (IL-3R positive) and NB4 (IL-3R negative) cells in acute myelocytic leukemia (AML) Methods HL-60 and NB4 cells were treated with different concentrations of IL-3-PE38KDEL. The proliferation inhibition was measured by MTT assay. The inhibitory rate and half-maximal inhibitory concentration (IC50) were calculated. The HL-60 and NB4 cells were divided into IL-3-PE38KDEL group and IL-3 + IL-3-PE38KDEL group, and IL-3 (100 ng / ml) After 4 hours, IL-3-PE38KDEL group and IL-3 + IL-3-PE38KDEL group were added with IC50 concentration of IL-3-PE38KDEL to calculate the inhibition rate of cell proliferation; flow cytometry was used to detect the cell cycle of the two kinds of AML cells Changes and apoptosis. Results IL-3-PE38KDEL inhibited the proliferation of HL-60 and NB4 cells in a concentration-dependent manner, and inhibited the proliferation of HL-60 cells more than NB4 cells (P <0.05). IC50 values were 2.5 μg / ml and 259.2 μg / ml. When the concentration of IL-3-PE38KDEL was IC50, the average inhibition rates of HL-60 and NB4 cell proliferation after IL-3 treatment were 32.20% and 49.00%, respectively. Most of HL-60 cells treated with IL-3-PE38KDEL were arrested in G1 / S phase. The average apoptotic rate in HL-60 cells (19.71%) was significantly higher than that in IL-3 + IL-3-PE38KDEL group (9.39%) (P <0.05) Significance (P> 0.05). Conclusion IL-3-PE38KDEL can inhibit the proliferation of HL-60 cells and induce the apoptosis of IL-3R-positive AML cells. However, IL-3-PE38KDEL has a weaker inhibitory effect on IL-3R-negative AML cells.