Polymyxin B (PMB), a last-line antibiotic used against antibiotic-resistant superbugs, causes undesirable cytotoxic side effects. However, its mechanisms remain unknown. In this study, we unexpectedly found that caspase-3, a main executor of apoptosis, plays a protective role in PMB-induced cytotoxicity. Caspase-3 knockout (KO) cells exhibited higher susceptibility to PMB-induced cytotoxicity compared with wild-type (WT) cells, accompanied by increased levels of reactive oxygen species (ROS). Interestingly, co-treatment with the antioxidant N-acetylcysteine (NAC) rescued cell viability to a similar extent as WT cells. Furthermore, PMB failed to facilitate the processing of inactive caspase-3 (pro-caspase-3) into active forms, suggesting that pro-caspase-3 nonenzymatically suppresses PMB-driven ROS accumulation and its cytotoxicity. Thus, our findings that demonstrate the potential ability of PMB to stimulate ROS generation, but which is normally masked by pro-caspase-3-dependent mechanisms, may provide novel insights into the mechanisms of PMB-induced side effects.
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This work was supported by JSPS KAKENHI Grant Numbers JP18H02567 and JP18K06622, and by MEXT KAKENHI JP17H05518 and JP19H05282. This work was also supported by the Fugaku Trust for Medicinal Research, the Takeda Science Foundation, and the Division for Interdisciplinary Advanced Research and Education (DIARE) Tohoku University.
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Yokosawa, T., Yamada, M., Noguchi, T. et al. Pro-caspase-3 protects cells from polymyxin B-induced cytotoxicity by preventing ROS accumulation. J Antibiot 72, 848–852 (2019). https://doi.org/10.1038/s41429-019-0216-6
The Journal of Toxicological Sciences (2020)