Mitomycin C Induces Apoptosis in Rheumatoid Arthritis Fibroblast-Like Synoviocytes via a Mitochondrial-Mediated Pathway
Mitomycin C Induces Apoptosis in Rheumatoid Arthritis Fibroblast-Like Synoviocytes via a Mitochondrial-Mediated Pathway
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Mitomycin C 通过线粒体介导的途径诱导类风湿性关节炎成纤维细胞样滑膜细胞凋亡。
DOI：
10.1159/000373938
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发表时间：
2015-01-01
影响因子：
--
通讯作者：
Cao, Xiaojian
中科院分区：
文献类型：
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作者：
Yan, Chuqi;Kong, Dechao;Cao, Xiaojian
背景/目的：类风湿关节炎(RA)是一种以滑膜细胞明显增生和成纤维细胞样滑膜细胞(FLS)的生长和死亡之间的潜在失衡为特征的全身性慢性炎症性疾病。丝裂霉素C(MMC)具有抑制成纤维细胞增殖和诱导成纤维细胞凋亡的作用。然而，MMC对人RA FLS增殖和凋亡的影响及其可能的作用机制尚不清楚。方法：采用细胞计数试剂盒-8法检测细胞存活率。Annexin V-FITC/PI双染和末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记分析细胞凋亡。用流式细胞仪检测细胞内活性氧(ROS)的产生，用荧光显微镜观察线粒体膜电位(ΔΨm)的变化。免疫印迹法检测细胞凋亡相关蛋白的表达。结果：MMC可显著降低RA FLS细胞存活率，诱导细胞凋亡。此外，与对照组相比，MMC暴露可刺激ROS的产生并扰乱ΔΨm。此外，MMC还可促进线粒体细胞色素c的释放、Bax/Bcl2的比值、caspase-9和caspase-3的激活以及随后的聚(ADP-核糖)聚合酶的切割。结论：MMC抑制RA FLS细胞增殖并诱导其凋亡，其机制可能与线粒体信号转导途径有关。
Background/Aims: Rheumatoid arthritis (RA) is a systemic chronic inflammatory disease characterised by prominent synoviocyte hyperplasia and a potential imbalance between the growth and death of fibroblast-like synoviocytes (FLS). Mitomycin C (MMC) has previously been demonstrated to inhibit fibroblast proliferation and to induce fibroblast apoptosis. However, the effects of MMC on the proliferation and apoptosis of human RA FLS and the potential mechanisms underlying its effects remain unknown. Methods: Cell viability was determined using the Cell Counting Kit-8 assay. Apoptotic cell death was analysed via Annexin V-FITC/PI double staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling. The production of intracellular reactive oxygen species (ROS) was assessed via flow cytometry, and the changes in mitochondrial membrane potential (ΔΨm) were visualized based on JC-1 staining via fluorescence microscopy. The expression of apoptosis-related proteins was determined via Western blot. Results: Treatment with MMC significantly reduced cell viability and induced apoptosis in RA FLS. Furthermore, MMC exposure was found to stimulate the production of ROS and to disrupt the ΔΨm compared to the control treatment. Moreover, MMC increased the release of mitochondrial cytochrome c, the ratio of Bax/Bcl-2, the activation of caspase-9 and caspase-3, and the subsequent cleavage of poly(ADP-ribose) polymerase. Conclusion: Our findings suggest that MMC inhibits cell proliferation and induces apoptosis in RA FLS, and the mechanism underlying this MMC-induced apoptosis may involve a mitochondrial signalling pathway.