Effect of intracellular calcium modulation on sulfur mustard cytotoxicity in cultured human neonatal keratinocytes.
Previous studies in human skin keratinocyte cultures have shown that sulfur mustard (HD) induces an immediate and irreversible increase in internal free calcium levels that was independent of external calcium concentrations. These findings suggested a role for calcium in the aetiology of HD-induced cell death and that modulation of intracellular calcium concentrations may assist in providing protection against this agent. In the current work, actively proliferating and confluent cultures of first passage neonatal human skin keratinocytes were used to assess the effect of altered intra- and extracellular calcium levels on HD toxicity. Treatment of cultures with the endoplasmic reticulum calcium ATPase inhibitor thapsigargin, or the calcium chelator BAPTA-AM, which reduce HD-induced elevation of intracellular free calcium, did not modulate the toxicity of HD. Furthermore, alteration of external calcium concentrations during these same experiments failed to elicit any change in the viability of HD-exposed cells. Treatment of confluent cultures with ionomycin at either low (100 microM) or high (1.2 mM) external calcium concentrations also failed to modulate the toxicity of HD in any way. It appears that in neonatal human skin keratinocytes in culture, HD-induced intracellular calcium perturbation does not play a major role in HD-induced cytotoxicity.
Sawyer
T W
TW
Therapy Group, Medical Countermeasures Section, Defence Research Establishment Suffield, Box 4000, Medicine Hat, Alberta, Canada. Thomas.Sawyer@dres.dnd.ca
Hamilton
M G
MG
D002118:Calcium / Q000378:metabolism*; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D015759:Ionomycin / Q000378:metabolism; D007476:Ionophores / Q000378:metabolism; D015603:Keratinocytes / Q000187:drug effects*; D009151:Mustard Gas / Q000633:toxicity*; D012867:Skin / Q000166:cytology; D019284:Thapsigargin / Q000494:pharmacology
D002614:Chelating Agents; D004791:Enzyme Inhibitors; D007476:Ionophores; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D015759:Ionomycin; D019284:Thapsigargin; D002118:Calcium; D009151:Mustard Gas