摘要
目的探讨雷公藤红素对人骨肉瘤细胞HOS的杀伤作用及其机制。方法将人骨肉瘤细胞HOS用各种不同浓度的雷公藤红素治疗后,采用MTT法检测雷公藤红素对HOS细胞活力的抑制作用;采用流式细胞术检测HOS细胞凋亡及活性氧簇(ROS)的产生情况;Western blot检测HOS细胞cleaved caspase-9、cleaved caspase-3和磷酸化JNK的表达水平。结果雷公藤红素对HOS细胞活力的抑制效应呈剂量依赖性,1、2、3、4、5μmol/L的雷公藤红素组细胞活力抑制率分别为(23.6±3.5)%、(43.7±5.4)%、(57.8±6.9)%、(75.5±6.4)%、(83.7±7.3)%,各雷公藤组细胞活力抑制力与对照组的零抑制力比较,差异均有统计学意义(P均<0.05);雷公藤红素可显著诱导HOS细胞发生凋亡,与对照组凋亡率(2.3±0.5)%比较,1、3μmol/L雷公藤红素组凋亡率分别为(17.2±2.2)%(P<0.05)、(39.5±3.6)%(P<0.05)。雷公藤红素可显著诱导HOS细胞ROS的产生,与对照组ROS阳性率(1.3±0.4)%比较,1、3μmol/L雷公藤红素组ROS阳性率分别为(13.6±1.5)%(P<0.05)、(29.4±3.3)%(P<0.05)。雷公藤红素处理后HOS细胞cleaved caspase-9、cleaved caspase-3和磷酸化JNK的表达水平均显著上升。结论雷公藤红素或通过ROS/JNK途径诱导人骨肉瘤细胞发生caspase依赖的凋亡。
Objective To explore the anti-tumor effect of celastrol on human osteosarcoma （HOS） cells and the underlying mechanism. Methods HOS cells were treated with various concentrations of celastrol, and then the cell viability was measured by using MTT assay. Apoptosis induction and ROS production were determined by flow eytometry. The expression of cleaved easpase-9, cleaved caspase-3 and phospho-JNK were evaluated by western blotting. Results The viability of HOS cells were significantly inhibited by eelastrol treatment in a dose-dependent manner, and the inhibition rate was as follows： 23.6%±3.5% in 1μmol/Lcelastrol group, 43.7%±5.4% in 2p, mol/L group, 57.8%±6.9% in 3μmol/Lgroup, 75.5%±6.4% in 4μmol/L group, 83.7%±7.3% in 5μmol/Lgroup （all P〈0.05）. Celastrol treatment significantly induced apoptosis in HOS cells, and the apoptotic rate was significantly different between control group and 1μmol/L and 3μmol/L eelastrol groups （2.3%±0.5% vs 17.2%±2.2% and 39.5%±3.6%, P〈0.05）. The ROS positive rate in HOS cells in 1μ mol/L and 3p, mol/L celastrol groups were significantly higher than that in control group induced（13.6%±1.5% and 29.4%±3.3% vs 1.3%±0.4%, P〈0.05）. Celastrol treatment significantly up-regulated the expression of cleaved caspase-9, cleaved caspase-3, and phospho-JNK in HOS cells. Conclusion Celastrol may induce caspase-dependent apoptotic cell death through ROS/JNKpathway in HOS cells.
出处
《浙江中西医结合杂志》
2015年第8期723-726,共4页
Zhejiang Journal of Integrated Traditional Chinese and Western Medicine
