U0126对急性白血病细胞HL-60增殖与凋亡的影响

Effect of U0126 on Proliferation and Apoptosis in Acute Leukemia Cell HL-60

目的:探讨U0126对急性白血病细胞HL-60增殖与凋亡的影响。方法:用不同浓度(0、5、10、20μmol/L)的U0126处理HL-60细胞24、48、72 h,MTT法检测细胞活力,Hoechst染色检测细胞形态,EdU染色检测细胞增殖,流式细胞术检测细胞周期,Western blot检测细胞中增殖细胞核抗原(PCNA)、生存素(Survivin)、p21和p27表达及细胞外调节蛋白激酶(ERK1/2)磷酸化水平。结果:5、10、20μmol/L的U0126处理HL-60细胞24、48、72 h,能显著提高细胞增殖抑制率(P<0.01),具有时间与剂量依赖性(r=0.421、0.478)。与0μmol/L比较,5、10、20μmol/L的U0126能使细胞核发白,致密浓染,细胞增殖率降低(P<0.01),细胞周期阻滞在G1期(P<0.01),PCNA、Survivin及p-ERK1/2表达均下调(P<0.01),p21与p27表达均上调(P<0.01)。结论:U0126能显著抑制白血病细胞HL-60增殖,诱导细胞凋亡。

Objective:To explore the effect of U0126 on proliferation and apoptosis in acute leukemia cell HL-60.Method:U0126 cell was cultured with 0,5,10,20μmol/L dose of U0126 for 24,48,72 h.Cell viability was measured by MTT assay.Cell morphology was detected by Hoechst stainning.Cell proliferation was detected by EdU staining.Cell cycle was measured by flow cytometry.The expression of Proliferating Cell Nuclear Antigen(PCNA),Survivin,p21,p27 and the level of extracellular regulated protein kinases(ERK1/2)phosphorylation were measured by Western blot.Result:HL-60 cell viability was reduced for treatment with 5,10,20μmol/L dose of U0126 for 24,48,72 h(P<0.01),time-and dose-dependent manner(r=0.421,0.478).5,10,20μmol/L dose of U0126 reduced cell proliferation rate(P<0.01),made cell nucleus dense and thick,made cell cycle arrested in the G1 phase(P<0.01),down-regulated the expression of PCNA,Survivin and p-ERK1/2(P<0.01),up-regulated the expression of p21 and p27(P<0.01).Conclusion:U0126 can significantly inhibit the proliferation of HL-60 cells and induce apoptosis.