DADS诱导人白血病细胞HL-60凋亡的流式细胞术检测

Detection of Human Leukemia HL-60 Cells Apoptosis Induced by Diallyl Disulfide With Flow Cytometry

目的运用流式细胞术检测二烯丙基二硫(diallyl disulfide,DADS)诱导人白血病HL-60细胞凋亡的作用。方法实验分对照组和DADS处理组,进行体外细胞培养,进行细胞形态学观察,应用流式细胞术检测凋亡率、早期凋亡细胞以及caspase-3活性。结果DADS诱导前后,HL-60细胞形态学发生明显改变,PI单染法检测结果:随着不同浓度DADS作用HL-60细胞24 h,HL-60细胞出现明显亚二倍峰,细胞周期也发生了相应的改变;Annexin V/PI双染法检测结果:10 mg/L DADS作用HL-60细胞4、8、12和24 h后,早期凋亡细胞显著增加,具有明显的时间依赖性(P<0.05);间接免疫荧光流式细胞术结果显示在DADS诱导HL-60细胞凋亡过程中有caspase-3的活化,且与作用时间呈依赖关系。结论流式细胞术检测结果表明,DADS能有效诱导HL-60细胞凋亡。

Objective Use flow cytometry to detect diallyl disulfide inducing human leukemia IlL -60 cells apoptosis. Methods HL -60 cells were treated with different concentrations of DADS. After incubation of HL -60 cells with DADS, the effects of DADS on HL - 60 cell growth inhibition were estimated by morphology, flow eytometry with PI and Annexin V/PI staining were used to determine apoptosis ceils and the expression of actived easpase - 3. Results After HL- 60 cells were exposed to DADS, the typical apoptotic morphological changes were observed under the light microscope, after treated with different concentrations of DADS for 24 h, HL - 60 cells presented obvious sub diploid peaks （ P 〈 0.05） and different cell cycle arrest . After treated with 10 mg/L DADS for 4, 8, 12, 24 h, early apoptotie cells were greatly increased （ P 〈 0.05 ） and the expression of active caspase - 3 was increased in a time - dependent manner. Conclusion Flow eytometry method show that DADS can efficiently induce human leukemia HL- 60 cells apoptosis.