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目的:研究槲皮素对白血病细胞的杀伤作用以及诱导的凋亡研究。方法:CCK-8试剂盒检测不同浓度的槲皮素(0~100μM)对白血病K562和HL-60细胞活性的影响;Hoechst33342染色技术观察40u M槲皮素处理细胞24和48小时后细胞核形态的变化;流式细胞仪检测细胞凋亡情况;caspase8、9、3(半胱天冬酶)特异性抑制剂预处理K562细胞后,检测药物对细胞活性的影响;40u M槲皮素处理细胞24和48小时后,Western blot检测细胞中caspase家族蛋白8、9、3的表达情况。结果:0~100u M的槲皮素能够引起K562和HL-60细胞剂量依赖型的活性下降,40u M的槲皮素处理细胞24和48小时后,K562细胞核明显发生皱缩,引起凋亡,随着处理时间增长,凋亡率增加,处理24和48小时后细胞的凋亡率分别为(45.8±2.06)%和(83.6±1.89)%。caspase 9、3抑制剂预处理K562细胞后,槲皮素能够引起细胞活性的增加,而caspase 8抑制剂预处理后,槲皮素没有引起细胞活性的上升。Western blot检测结果表明40μM的槲皮素处理细胞后能够上调caspase 9、3蛋白的表达,而没有引起caspase 8蛋白表达量的上升,随着处理时间的增长,caspase 9、3的表达量更高。结论:槲皮素对白血病细胞具有很强的杀伤作用,并诱导细胞caspase 9、3蛋白参与的内源性通路介导的凋亡。
Objective: To study the killing effect of quercetin on leukemia cells and its apoptosis. Methods: The effects of different concentrations of quercetin (0-100 μM) on the activity of leukemia K562 and HL-60 cells were detected by CCK-8 kit. The morphological changes of nuclei were observed by Hoechst33342 staining at 24 and 48 hours after treated with 40 uM quercetin The apoptosis of K562 cells was detected by flow cytometry (FCM). After K562 cells were pretreated with caspase8,9,3 (caspase-specific inhibitor), the effects of drugs on cell viability were measured. The cells treated with 40u M quercetin After 48 hours, Western blot was used to detect the expression of caspase family protein 8, 9 and 3 in the cells. Results: Quercetin 0-100u M induced a dose-dependent decrease in K562 and HL-60 cells. After treated with 40u M quercetin for 24 and 48 hours, K562 cells obviously shrunk and induced apoptosis, With the increase of treatment time, the apoptosis rate was increased, and the apoptotic rate was (45.8 ± 2.06)% and (83.6 ± 1.89)% after 24 and 48 hours treatment, respectively. Quercetin increased cell viability after K562 cells were pretreated with caspase 9,3 inhibitor, whereas quercetin did not induce cell viability after pretreatment with caspase 8 inhibitor. Western blot results showed that 40μM quercetin could up-regulate the expression of caspase 9 and 3 but not caspase 8, and the expression of caspase 9 and 3 increased with the increase of treatment time . CONCLUSIONS: Quercetin has a strong killing effect on leukemic cells and induces endogenous pathway-mediated apoptosis in which caspase 9 and 3 proteins are involved.