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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 investigate the effects of 17-hydroxy-petridrel lactone B (HJB) on the proliferation and apoptosis of human leukemia K562 cells. Methods: K562 cells were divided into three groups: blank control group, 5-fluorouracil group (concentration 100μmol.L-1), HJB group (6.25,12.5,25,50,100,200,400μmol.L-1 ). The cells were treated with different concentrations of HJB for 24 h and 200 μmol·L-1 for different time (12,24,48 h) and MTT assay, and compared with blank control group and 5-fluorouracil group. Flow cytometry was used to detect apoptosis rate by Annexin V-FITC / PI. Caspase-3 and caspase-8 were detected by spectrophotometry Caspase-8) relative activity. Results: Compared with the blank control group, HJB significantly inhibited the proliferation of K562 cells in a dose-dependent and time-dependent manner (P <0.05). IC50 was 158.7 μmol.L-1 after 24 h. HJB could induce the apoptosis of K562 cells. The apoptosis rate of K562 cells treated with 50,100 and 200μmol.L-1 for 24 h were significantly higher than that of the blank group (P <0.05), and the concentration of Caspase-3 and Caspase8 (P <0.05) compared with blank control group, and showed a concentration-dependent relationship. Conclusion: HJB significantly inhibits the growth of K562 cells and induces its apoptosis in vitro, and the death receptor pathway may be a mechanism of inducing apoptosis.