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目的探讨异硫氰酸苄酯(benzyl isothiocyanate,BITC)对人脑胶质瘤细胞系U-87 MG的活性氧(ROS)的诱导作用及其机制。方法应用MTS法检测BITC对肿瘤细胞生长的影响,2和5μmol·L-1 BITC作用U-87 MG细胞后,应用流式细胞术检测肿瘤细胞内活性氧(ROS)含量的变化,生化法检测GSH以及氧化应激相关的线粒体呼吸链复合体Ⅲ、过氧化物歧化酶(SOD)和醌还原酶(quinone reductase,QR)的活性变化,Western blotting法和报告基因技术检测p38-MAPK和相关转录因子ARE的转录活性变化。结果 BITC对脑胶质瘤细胞U-87 MG具有明显的抑制作用,其IC50值为15.2μmol·L-1,2和5μmol·L-1 BITC作用肿瘤细胞24 h后,ROS产生分别为对照组的376.3%和607.5%(P<0.05),GSH水平分别为对照组的71.3%和44.9%(P<0.05),SOD活性分别为对照组的63.5%和21.8%(P<0.05),QR活性分别为对照组的55.2%和26.7%(P<0.05),呼吸链复合体III活性分别为对照组的48.5%和37.6%(P<0.05),p38-MAPK的磷酸化水平显著上升,ARE的转录活性分别为对照组的141.1%和215.2%(P<0.05)。结论 BITC可诱导脑肿瘤细胞U-87 MG中ROS产生,可能与调节胞内的氧化应激相关基因表达有关。
Objective To investigate the effect and mechanism of benzyl isothiocyanate (BITC) on reactive oxygen species (ROS) in human glioma cell line U-87 MG. Methods The effect of BITC on the growth of tumor cells was detected by MTS method. After treated with 2 and 5 μmol·L -1 BITC, the changes of reactive oxygen species (ROS) in tumor cells were detected by flow cytometry. GSH and mitochondrial respiratory chain complex Ⅲ, the activities of superoxide dismutase (SOD) and quinone reductase (QR), and the changes of p38-MAPK and related transcription by Western blotting and reporter gene technology Transcriptional activity of factor ARE changes. Results BITC had a significant inhibitory effect on glioma U-87 MG cells with IC50 values of 15.2μmol·L -1 and 5μmol·L -1 BITC for 24 h, (P <0.05). The levels of GSH were 71.3% and 44.9% (P <0.05) in the control group and 63.5% and 21.8% (P < (P <0.05). The activity of respiratory chain complex III was 48.5% and 37.6% (P <0.05) respectively in the control group, the phosphorylation level of p38-MAPK was significantly increased, the expression of ARE The transcriptional activities were 141.1% and 215.2% in the control group (P <0.05). Conclusion BITC induces ROS production in U-87 MG cells, which may be related to the regulation of intracellular oxidative stress-related gene expression.