Mitomycin C Induces Apoptosis in Rheumatoid Arthritis Fibroblast-Like Synoviocytes via a Mitochondrial-Mediated Pathway

Abstract

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.

miR-338-5p Regulates the Viability, Proliferation, Apoptosis and Migration of Rheumatoid Arthritis Fibroblast-Like Synoviocytes by Targeting NFAT5

“…With further efforts, Ogando and his collogues identified miR-223 as a negative regulator of AHR/ARNT pathway by targeting ARNT, which negatively modulates the downstream Notch effectors [32]. Yan C et al identified mitomycin C induces apoptosis in rheumatoid arthritis fibroblast- Cellular Physiology and Biochemistry Cellular Physiology and Biochemistry like synoviocytes [33]. In our study, the expression level of miR-338-5p in RA synovium was significantly higher than that in normal synovium.…”

Effects of sesamin on primary human synovial fibroblasts and SW982 cell line induced by tumor necrosis factor-alpha as a synovitis-like model

“…Thus, in vitro models that use human cells were developed to study the mechanism of RA and possible therapeutic approaches [20]. Therefore, primary human fibroblast-like synoviocytes from RA patients have been used to screen and study the unique effects of many drugs and phytochemicals [21–23]. Despite the advantages of primary human fibroblast-like synoviocytes, they present certain inconveniences; for instance, primary mammalian cells have a limited replicable lifespan and ultimately enter a state of senescence in which they remain metabolically active but fail to proliferate.…”

“…Thus, primary human fibroblast-like synoviocytes isolated from RA patients have been used to study the effects of a variety of drugs and phytochemicals [21–23]. However, there are some difficulties in using RA-derived synovial fibroblasts.…”

Effect of 1,25-(OH)2D3 on Proliferation of Fibroblast-Like Synoviocytes and Expressions of Pro-Inflammatory Cytokines through Regulating MicroRNA-22 in a Rat Model of Rheumatoid Arthritis

“…RA FLS can exhibit resistance to apoptosis caused by apoptotic stimuli, further facilitating FLS hyperplastic growth and the destruction of articular cartilage [22]. Therefore, inhibiting the proliferation of RA FLS and prpmoting apoptosis of FLS are therapeutic approaches for the treatment of RA.…”