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目的探讨阿糖胞苷诱导人MDS细胞株MUTZ-8细胞凋亡的可能机制。方法将细胞分为以下5组:阴性对照组、0.1μmol/L Ara-c组、1μmol/L Ara-c组、10μmol/L Ara-c组、10μmol/L Ara-c+62.5μmol/L ALA组。用MTT法测定阿糖胞苷对各组细胞抑制率的影响,用Annexin V-FITC/PI双染法及Hoechst33258荧光染色观察阿糖胞苷对各组细胞凋亡的影响,并用流式细胞仪检测各组细胞线粒体膜电位的变化。运用二氢二氯荧光素(DCFH-DA)为细胞内活性氧探针,通过荧光显微镜及流式细胞仪检测各组细胞内活性氧水平的变化。结果 MTT结果提示,阿糖胞苷处理组细胞增殖抑制率高于阴性对照组,二者相比差异具有统计学意义(P<0.05)。流式细胞仪结果提示,不同浓度的阿糖胞苷作用MUTZ-8细胞后,凋亡率明显升高,线粒体膜电位明显下降,与阴性对照组相比,差异具有统计学意义(P<0.05)。流式细胞仪及荧光显微镜检测均显示,阿糖胞苷作用后细胞内活性氧(reactive oxygen species,ROS)明显上升。而加入抗氧化剂α-硫辛酸后细胞内ROS表达水平下降。结论阿糖胞苷明显抑制了人MDS细胞株MUTZ-8的生长并诱导其凋亡,可能与阿糖胞苷影响了细胞内ROS水平有关。
Objective To investigate the possible mechanism of cytarabine-induced apoptosis in human MDS cell line MUTZ-8. Methods The cells were divided into the following 5 groups: negative control group, 0.1μmol / L Ara-c group, 1μmol / L Ara-c group, 10μmol / L Ara-c group, 10μmol / L Ara-c + 62.5μmol / group. The effect of cytarabine on the cell inhibition rate was determined by MTT assay. The effect of cytarabine on the apoptosis of each group was observed by Annexin V-FITC / PI double staining and Hoechst33258 fluorescence staining. The change of mitochondrial membrane potential in each group was detected. The intracellular reactive oxygen species (ROS) levels in each group were detected by fluorescence microscopy and flow cytometry with DCFH-DA as an intracellular reactive oxygen species (ROS) probe. Results The results of MTT suggested that the cell proliferation inhibition rate of cytarabine group was higher than that of negative control group, the difference was statistically significant (P <0.05). The results of flow cytometry showed that the apoptosis rate of MUTZ-8 cells was significantly increased and the mitochondrial membrane potential was significantly decreased after treated with different concentrations of cytarabine, which was significantly lower than that of the negative control group (P <0.05 ). Flow cytometry and fluorescence microscopy showed that cytosolic oxygen species (ROS) increased significantly after cytarabine treatment. However, the level of intracellular ROS decreased after adding anti-oxidant α-lipoic acid. Conclusion Cytarabine significantly inhibits the growth of human MDS cell line MUTZ-8 and induces its apoptosis, which may be related to the effect of cytarabine on the intracellular ROS level.