二烯丙基二硫诱导人白血病K562细胞凋亡及其对Fas、FasL及caspase-8表达的影响

Effects of diallyl disulfide on apoptosis of human leukemia K562 cells and expression of Fas, FasL and caspase-8

摘要:

目的：探讨二烯丙基二硫（DADS）诱导白血病 K562 细胞凋亡的作用及其机制。方法：采用 Hoechst 33258 染色法观察细胞凋亡形态学变化；流式细胞术分析不同浓度及不同作用时间 DADS 对 K562 细胞凋亡的诱导作用；RT-PCR法检测DADS作用48 h后 Fas、FasL、caspase-8 mRNA的表达。结果：DADS可诱导K562 细胞凋亡。DADS(作用细胞24 h)浓度从10 mg/L增至40 mg/L时，其对K562细胞的凋亡率由（11.60±0.83）%增至（37.94±0.87）%；DADS(40 mg/L浓度组)作用时间从24 h增至72 h，其对K562细胞的凋亡率由（37.94±0.87）% 增至（47.02±0.66）%，各实验组随时间、浓度增加凋亡率明显增加 (P<0.05)。DADS作用48 h后，Fas、caspase-8 mRNA 表达水平较对照组上调；FasL mRNA 较对照组下调（P＜0.05）。结论：DADS 呈时间和浓度依赖性诱导白血病K562 细胞凋亡，其凋亡机制可能与上调 Fas、caspase-8，下调 FasL有关。

Abstract:

OBJECTIVE: To study the effects of diallyl disulfide (DADS) on apoptosis of human leukemia K562 cells and possible mechanisms. METHODS: The morphologic changes of leukemia K562 cells after DADS treatment were observed by Hoechst 33258 staining. Cell apoptosis rates after different concentrations and different durations of DADS treatment were determined by flow cytometry. Fas, FasL and caspase-8 mRNA expression was estimated by reverse transcription-polymerase chain reaction (RT-PCR) 48 hrs after DADS treatment. RESULTS: The characteristics of apoptosis in K562 cells induced by DADS were observed. After 24 hrs of DADS treatment, the apoptosis rate of K562 cells increased from (11.60±0.83)% at the concentration of 10 mg/L to (37.94±0.87)% at the concentration of 40 mg/L. The apoptosis rate of K562 cells increased after 40 mg/L DADS with the increasing time from (37.94±0.87)% (24 hrs) to (47.02±0.66)% (72 hrs). Expression of Fas and caspase-8 mRNA increased, while FasL mRNA expression decreased significantly 48 hrs after DADS treatment compared with the control group (P<0.05). CONCLUSIONS: DADS can induce apoptosis of human leukemia K562 cells in a time- and concentration-dependent manner, possibly through increasing Fas and caspase-8 expression and decreasing FasL expression.