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

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100μmol.L-1),HJB组(浓度为6.25,12.5,25,50,100,200,400μmol.L-1)。用不同浓度的药物作用24 h和200μmol.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158.7μmol.L-1。HJB可诱导K562细胞凋亡,用50,100,200μmol.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。

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＆lt;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＆lt;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.