紫草素衍生物SK36对白血病HL-60细胞增殖及凋亡作用的影响

The effects of shikonin derivative SK36 on the proliferation and apoptosis of HL-60 cells

目的:探讨紫草素衍生物5,8-二甲基-2-β-羟基-异戊酰紫草素(5,8-dimethyl-2-β-hydroxy-isovaleryl shikonin,SK36)对人白血病细胞株HL-60增殖及凋亡的影响,并初步探讨其作用机制。方法:CCK-8法检测SK36对HL-60细胞的增殖抑制作用;光学显微镜下观察细胞形态的变化;AnnexinⅤ/PI双标记法检测细胞凋亡率;FCM法检测caspase-3活性;JC-1染色法检测细胞线粒体跨膜电位。结果:SK36能明显抑制HL-60细胞的增殖,呈剂量和时间依赖性;在光学显微镜下可见典型的细胞凋亡形态学变化;FCM法检测结果显示,SK36作用HL-60细胞24和48 h时的细胞凋亡率分别为(55.55±2.88)%和(72.12±14.50)%,明显高于对照组;caspase-3活性增加,线粒体跨膜电位发生崩塌,且随着时间的延长其效应更加明显,与对照组相比差异有统计学意义(P<0.05)。结论:SK36在体外可抑制HL-60细胞增殖并诱导其凋亡。

Objective：To investigate the effects of shikonin derivatives 5,8-dimethyl-2-β-hydroxy-isovaleryl shikonin（SK36） on proliferation and apoptosis of human leukemia cell line HL-60 in vitro and elucidate its mechanism of action.Methods：The inhibitory effects of SK36 on proliferation of HL-60 cells were detected using CCK-8 assay.Cell morphological alteration was observed by using light microscopy.Cell apoptosis was analyzed by flow cytometry（FCM） with AnnexinⅤ/PI double labeling method.Caspase-3 activity was measured with FCM;mitochondial transmembrance potential was analyzed by JC-1 staining.Results：SK36 greatly inhibited the proliferation of HL-60 cells in a concentration-and time-dependent manner.After being treated with SK36,the typical apoptotic morphological changes were observed in HL-60 cells under the light microscope.The cell apoptotic rate of HL-60 cells evaluated by FCM with AnnexinⅤ/PI double labeling staining were（55.55±2.88）% and（72.12±14.50）% at 24 and 48 h after treatment,respectively,which were significantly higher than that of the control group.There was a significant increase in caspase-3 activity and mitochondrial transmembrane potential collapse compared with the control group（P0.05）.The effect was more potent with the prolongation of treatment time（P0.05）.Conclusion：SK36 inhibits the proliferation and induces apoptosis of HL-60 cells in vitro.