17-羟-岩大戟内酯B对K562细胞增殖及凋亡的影响

Effect of 17-hydroxy-jolkinolide B on Proliferation and Apoptosis of K562 Cells

摘要:

目的: 探讨17-羟-岩大戟内酯B(HJB)对人白血病K562细胞增殖及凋亡的影响。 方法: 将K562细胞分为3组,分别为:空白对照组,5-氟脲嘧啶组(浓度为100 μmol·L^-1),HJB组(浓度为6.25, 12.5,25,50,100,200,400 μmol·L^-1)。用不同浓度的药物作用24 h和200 μmol·L^-1的HJB浓度作用不同时间(12, 24, 48 h),四甲基偶氮唑蓝(MTT)法检测细胞活性,并与空白对照组和5-氟脲嘧啶组作对比;流式细胞仪Annexin V-FITC/PI检测细胞凋亡率;分光光度法检测半胱氨酸蛋白酶-3(Caspase-3)和半胱氨酸蛋白酶-8(Caspase-8)的相对活性。 结果: 与空白对照组相比,HJB对K562细胞的增殖有显著性抑制作用,并呈浓度依赖关系及时间依赖关系(P<0.05),作用24 h后IC₅₀为158.7 μmol·L^-1。HJB可诱导K562细胞凋亡,用50, 100, 200 μmol·L^-1药物分别处理细胞24 h后,早期凋亡率较空白组明显升高(P<0.05),且呈浓度依赖关系;Caspase-3及Caspase8的相对活性均较空白对照组升高(P<0.05),并呈浓度依赖关系。 结论: HJB明显抑制体外K562细胞生长并诱导其发生凋亡,且死亡受体途径可能是诱导其凋亡的一个机制。

Abstract:

Objective: To study the effects of proliferation and apoptosis on K562 cells induced by 17-hydroxy-jolkinolide B (HJB). Method: After the cells were treated with HJB at various concentrations (0, 6.25, 12.5,25,50,100,200,400 μmol·L^-1)for 24 h, or at 200 μmol·L^-1for various times (12, 24, 48 h), cell viability was measured by 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT). Annexin V-FITC was used to test the apoptosis rate. Absorption spectrometry was adopt to measure the relative activity of Caspase-3 and Caspase-8. Result: The HJB evidently inhibited the growth of K562 cells in a time and dose-dependent manner(P<0.05), with IC₅₀value for 24 h being 158.7 μmol·L^-1, and it could induce apoptosis of K562 cells. After HJB treatment at 50, 100, 200 μmol·L^-1 for 24 h, the apoptosis rate of K562 cells was 12.14%, 19.26% and 21.78%, respectively(P<0.05),and the relative activity of Caspase-3 or Caspase-8 was advanced with dose-dependent manner. Conclusion: The HJB evidently inhibits the growth of K562 and induces the apoptosis of K562.The mechanism of apoptosis-induced of HJB may be mediated by death receptor pathways.