Differentiation of HL-60 cells inducted by Herba Saginae Japonicae ethanol extract
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摘要: 目的 探讨漆姑草醇提物(HSJ)对人早幼粒白血病细胞(HL-60)的诱导分化作用。方法 实验设漆姑草250.0、125.0、62.5 μg/mL组、阳性对照组(吡柔比星100 μg/mL)、对照组;噻唑蓝(MTT)法检测HL-60细胞增殖抑制率;瑞氏-吉姆萨染色法观察细胞形态;硝基四氮唑蓝(NBT)还原实验法测定细胞分化能力;流式细胞术检测分化相关抗原CD11b、CD14阳性细胞的表达;Western blot法检测c-myc蛋白表达。结果 与对照组比较,漆姑草组HL-60细胞核浆比例减小,核仁变为肾形或蚕豆形,出现明显的细胞分化形态,细胞增殖受到不同程度抑制;对照组HL-60细胞NBT还原率为(0.83±0.29)%,CD11b和CD14阳性细胞表达率分别为(41.13±0.42)%、(53.30±7.23)%,c-myc蛋白表达量为(0.71±0.02);漆姑草250.0、125.0、62.5 μg/mL组HL-60细胞NBT还原率[分别为(16.67±1.76)%、(10.83±1.26)%、(6.00±0.87)%]升高,CD11b和CD14阳性细胞表达率[分别为(66.77±4.27)%、(52.27±0.95)%、(46.27±0.38)%和(96.63±0.23)%、(85.25±3.80)%、(63.84±0.78)%]增加,c-myc蛋白表达量[分别为(0.38±0.01)、(0.45±0.01)、(0.58±0.02)]降低,差异均有统计学意义(均P<0.05)。结论 漆姑草醇提物可诱导HL-60细胞向成熟粒细胞、单核细胞方向分化。Abstract: Objective To study the effect of Herba Saginae Japonicae ethanol extract (HSJ) on the differentiation of human promyelocytic leukemia (HL-60) cells.Methods After 24 hours' incubation during logarithmic growth phase,the HL-60 cells were assigned into three experimental groups (treated with 250,125,62.5μg/mL HSJ),a positive control group (with 100 μg/mL pirarubicin),and a negative control group.The proliferation of HL-60 cells was detected with methyl thiazolyl tetrazolium bromide (MTT) assay;the cell morphology was observed with Wright-Giemsa staining; the cell differentiation ability was determined with nitroblue tetrazolium (NBT) reduction assay; the expressions of differentiation related antigen (CD11b and CD14) were measured with flow cytometry (FCM);and the expression of c-myc protein was detected with Western blot.Results Compared with the control group,the HSJ-treated HL-60 cells demonstrated apparent differentiation and inhibited proliferation,with decreased nucleus and cytoplasm ratios and kidney-or broad bean-shaped nucleoli.For the HSJ cells treated with 250,125,and 62.5 μg/mL HSJ,the NBT reduction rates were 16.67±1.76%,10.83±1.26%,and 6.00±0.87%;the CD11b positive rates were 66.77±4.27%,52.27±0.95%,and 46.27±0.38%; and the CD14 positive rates were 96.63±0.23%,85.25±3.80%,and 63.84±0.78%,respectively; all the indicators were significantly increased compared to those of negative control HSJ cells (0.83±0.29%,41.13±0.42%,and 53.30±7.23%)(P<0.05 for all).Wheaeas,the c-myc protein expressions (0.38±0.01,0.45±0.01,and 0.58±0.02) of the HL-60 cells treated with different dosages of HSJ decreased significantly compared to that (0.71±0.02) of the negative control HSJ cells (P<0.05).Conclusion HSJ can induce HL-60 cells differentiating into mature granulocytes or monocytes.
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Key words:
- Herba Saginae Japonicae /
- HL-60 cell /
- induced differentiation
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