灯盏花乙素对急性髓系白血病细胞增殖的影响及作用机制研究

Effect of Scutellarin on Proliferation of Acute Myeloid Leukemia Cells and Its Related Mechanism

目的:探讨灯盏花乙素(SCU)对急性髓系白血病(AML)细胞增殖、周期与凋亡的影响及相关分子机制。方法:体外培养人AML HL-60细胞,分别以0、2、4、8、16、32、64μmol/L浓度的SCU处理HL-60细胞,采用CCK-8法检测细胞增殖抑制率。以4、8、16μmol/L浓度的SCU处理HL-60细胞,并设置阴性对照组(NC组),采用流式细胞术检测细胞周期分布和细胞凋亡,Western blot法检测细胞周期、凋亡相关蛋白及JAK2/STAT3通路相关蛋白的表达。结果:SCU对HL-60细胞具有明显的增殖抑制作用,且增殖抑制作用呈浓度(r=0.958)、时间(r=0.971)依赖性。与NC组比较,SCU 4、8、16μmol/L组HL-60细胞G0/G1期比例、细胞凋亡率均显著升高,S期比例均显著降低(P<0.05);p21、p53、caspase-3、Bax蛋白相对表达量均显著升高,CDK2、cyclin E、Bcl-2蛋白相对表达量均显著降低(P<0.05),p-JAK2/JAK2、p-STAT3/STAT3比值均显著降低(P<0.05),且上述指标变化均呈浓度依赖性。结论:SCU可抑制AML细胞增殖,诱导细胞周期阻滞和细胞凋亡,其作用机制可能与调控JAK2/STAT3信号通路相关。

Objective:To investigate the effect of scutellarin(SCU)on proliferation,cell cycle and apoptosis of acute myeloid leukemia(AML)cells and its related molecular mechanism.Methods:Human AML HL-60 cells were cultured in vitro.The cells were treated with SCU at the concentration of 0,2,4,8,16,32,64μmol/L,and the inhibition rate of cell proliferation was detected by CCK-8 method.Then HL-60 cells were treated with SCU at the concentration of 4,8,16μmol/L,and the negative control group(NC group)was set.The cell cycle distribution and apoptosis were detected by flow cytometry,and the expression of cell cycle,apoptosis and JAK2/STAT3 pathway related proteins were detected by Western blot.Results:SCU significantly inhibited the proliferation of HL-60 cells in a concentration-and time-dependent manner(r=0.958,r=0.971).Compared with NC group,the proportion of cells in G0/G1 phase and apoptosis rate of HL-60 cells in 4,8,16μmol/L SCU group were significantly increased,and the proportion of cells in S phase was significantly decreased(P<0.05).The relative protein expression levels of p21,p53,caspase-3 and Bax were significantly increased,while the relative protein expression levels of CDK2,cyclin E and Bcl-2 were significantly decreased(P<0.05).The ratio of p-JAK2/JAK2 and p-STAT3/STAT3 were significantly decreased(P<0.05).The changes of above-mentioned indexes were concentration dependent.Conclusion:SCU can inhibit the proliferation of AML cells,induce cell cycle arrest and apoptosis,and its mechanism may be related to the regulation of JAK2/STAT3 signaling pathway.