Zinc-metallothionein genoprotective effect is independent of the glutathione depletion in HaCaT keratinocytes after solar light irradiation

Abstract

UV radiations are the major environmental factors that induce DNA damage of skin cells either by direct absorption (UVB), or after inducing an oxidative stress (UVA and UVB). Cells maintain a reducing intracellular environment to avoid genomic damage. MTs have been expected not only to control metal homeostasis but also counteract the glutathione (GSH) depletion induced by oxidative stress because of their high thiol content. Induction and redistribution of MTs in cultured human keratinocytes (HaCaT) in response to SSL, is an important cellular defense mechanism against DNA damage. Reduced glutathione (GSH) is another way of cellular protection against UV-induced oxidative stress. This study which extend our previous finding focused on the relation between intracellular GSH and Zn genoprotective effects after solar irradiation. HaCaT cells, depleted or not in GSH by a chemical treatment were used to compare MTs induction by Northern blot, expression by Western blot and localization using immunocytochemistry. Zn genoprotection experiments after SSL irradiation was carried out by the comet assay. We demonstrated that in absence of GSH, Zn-MTs could protect DNA after SSL irradiation and that GSH depletion has no effect on MTs induction and localization. Nuclear Zn-MTs could be responsible for this observed genoprotection in GSH depleted cells. So the GSH/Zn and the MT/Zn systems could be two independent but interacting mechanisms of cellular protection against SSL injury. © 2004 Wiley-Liss, Inc.