Effect of sulphur mustard on human skin cell lines with differential agent sensitivity

Abstract

The ability of sulphur mustard (HD) to induce DNA damage places limits on the efficacy of approaches aimed at protecting human cells from the cytotoxic effects of HD using a variety of protective agents such as thiol-containing esters and protease inhibitors. In the present study, potential alternative strategies were investigated by examining the differential effects of HD on G361, SVK14, HaCaT and NCTC 2544 human skin cells. The G361 cell line was more resistant to the cytotoxic effects of HD than the NCTC, HaCaT and SVK14 cell lines at HD doses of >3 and <100 microM HD as determined by the MTT assay. At 72 h after exposure to 60 microM HD there was up to an 8.8-fold difference (P < 0.0001) between G361 and SVK14 cell culture viability. Buthionine sulphoximine (BSO) pretreatment increased the sensitivity of all four cell lines to HD. A substantial proportion of the resistance of G361 cells to HD was attributable to BSO-mediated effects on antioxidant-mediated metabolism, although G361 cultures still retained a high degree of viability at 30 microM HD following BSO pretreatment. Cell cycle analysis confirmed that SVK14 cells were relatively more sensitive to HD, as shown by the 2.1-fold reduction (P < 0.0001) in the percentage of cells in G0/G1 phase 24 h after HD exposure compared with control cultures. This compared well with a 1.2-fold increase (P < 0.05) in the percentage of G361 cells in G0/G1 phase following HD exposure, suggesting the existence of a more efficient G0/G1 checkpoint control mechanism in this cell line. Manipulation of the cell cycle using various modulating agents did not increase the resistance of cell lines to the cytotoxic effects of HD.

Protective Effect of Liposome-Encapsulated Glutathione in a Human Epidermal Model Exposed to a Mustard Gas Analog

“…In the EpiDerm model, GSH liposomes showed significant protective effect even when applied 1 hour after CEES exposure. This observation further confirmed previous in vitro studies of CEES/HD toxicity, which have demonstrated that the antioxidants that reduce oxidative stress and prevent thiol depletion also attenuate CEES toxicity, whereas oxidants (hydrogen peroxide) [4] and agents that deplete intracellular glutathione (buthionine sulphoximine) [6, 42] sensitize human cell lines, in particular HaCaT [42], to CEES/HD toxicity. Thus, our study provides additional evidence of the critical role of oxidative stress in the pathogenesis of CEES/HD-induced injury in the skin.…”

“…However, this direct detoxification is possible only during a short time after the exposure, and other nucleophilic scavengers, for example, thiopurines [23] or exogenous peroxidases [44], are more effective in degrading of HD or CEES. Nevertheless, antioxidants and antioxidant liposomes have shown a great potential in preventive treatment of CEES/HD toxicity [3–5, 9, 10, 20, 21, 26, 34, 42, 45–48]. In addition, it is well known that mammalian skin cells exposed to CEES/HD release proinflammatory cytokines [27, 28, 37, 49] and other immune stimulators [50].…”

Efficacy of Glutathione in Ameliorating Sulfur Mustard Analog-Induced Toxicity in Cultured Skin Epidermal Cells and in SKH-1 Mouse Skin In Vivo

“…This finding suggests that the protective effect of NAC against CEES-caused toxicity occurs in part through an increase in GSH levels, and the other mechanisms could be related to its activity as a strong ROS scavenger; NAC is also known to act via the nuclear factor-B pathway and inhibit prostaglandin synthesis apart from being a GSH precursor (Atkins et al, 2000;Arfsten et al, 2007). CEES-induced depletion of GSH and generation of ROS have been shown in human lymphocytes that induce cell death and in human skin cells where treatment with GSHdepleting agent BSO showed an increased toxicity to HD (Han et al, 2004;Simpson and Lindsay, 2005). In the present study, we also observed that BSO pretreatment to both the skin epidermal cells caused an increase in CEES-caused toxicity.…”

2,6-Dithiopurine Blocks Toxicity and Mutagenesis in Human Skin Cells Exposed to Sulfur Mustard Analogues, 2-Chloroethyl Ethyl Sulfide and 2-Chloroethyl Methyl Sulfide

“…NCTC2544 cells are known to have inducible Phase I and Phase II detoxication activities (20, 21) and have previously been used for studies of sulfur mustard toxicity (22). The medium used for exposure to the alkyl mustards was modified to accommodate relatively high concentrations of the potential scavenger, DTP, which is not readily soluble in water, phosphate buffered saline or DMEM.…”