In vitro stress response induced by sulfur mustard in lung fibroblasts NHLF and human pulmonary epithelial cells A-549
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IN VITRO SYSTEMS
In vitro stress response induced by sulfur mustard in lung fibroblasts NHLF and human pulmonary epithelial cells A‑549 Petr Jost1 · Lubica Muckova1 · Jaroslav Pejchal1 Received: 28 April 2020 / Accepted: 9 July 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Sulfur mustard [bis(2-chloroethyl) sulfide; SM] is a highly poisonous chemical warfare agent. The mechanism of its cytotoxicity affects several pathways, which cause cell damage or death. The main organ affected in case of exposure to both aerosol and vapor is lungs. The present study focuses on time- and concentration-dependent changes in human lung fibroblasts NHLF and lung epithelial cell line A-549. The cells were treated with SM at the concentrations of 5, 10 and 100 µM and signs of stress response were evaluated during 1–72 h post-treatment. Parameters for testing included cell viability and morphology, loss of transmembrane mitochondrial potential, apoptosis, oxidative stress, changes in the cell cycle, and ATM kinase activation. The cytotoxic effect of SM resulted in a time-dependent decrease in viability of A-459 associated with apoptosis more markedly than in NHLF. We did not observe any generation of reactive oxygen species by SM. SM at concentrations of 5 and 10 µM induced the S-phase cell cycle arrest at both cell lines. On the other hand, 100 µM caused nonspecific cell cycle arrest. ATM kinase was activated transiently. The results indicate that NHLF cells are less prone to toxic damage by SM in case of cell viability, apoptosis and loss of transmembrane mitochondrial potential. The analysis provides a time-related cytotoxic profile of A-549 and NHLF cells for further investigation into the prevention of SM toxic effects and their potential treatment. Article Highlights (1) Sulfur mustard induced the time- and concentration-dependent cytotoxic effects on both A-549 cells and lung fibroblast NHLF. (2) Sulfur mustard at concentrations of 5 and 10 μM caused cell cycle arrest in S-phase, whereas 100 μM inhibited cell cycle completely. (3) Loss of mitochondria potential occurred with the latency of several hours. (4) SM induced transient activation of ATM kinase. (5) The DCFH-DA fluorescent probe did not confirm the generation of ROS. Keywords Sulfur mustard · Cytotoxicity · Apoptosis · Cell cycle · S-phase arrest · A-549 cell line · Lung fibroblasts NHLF
Introduction Sulfur mustard [SM; bis(2-chloroethyl) sulphide] is a cytotoxic and cytostatic agent with a blistering effect on human skin. The respiratory tract is a vital target organ as well as * Petr Jost [email protected] 1
Department of Toxicology and Military Pharmacy, Faculty of Military Health Sciences, University of Defence in Brno, Trebesska 1575, 500 01 Hradec Kralove, Czech Republic
the route of intoxication. Inhalation exposure to SM is often lethal or it could become a source of chronic symptoms and disability in long-term survivors (Seagrave et al. 2010; Weinberger et al. 2011). Elucidation of the mechanism of SM toxicity has, the
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