title: Differential responses of S100A2 to oxidative stress and increased intracellular calcium in normal, immortalized, and malignant human keratinocytes.
abstract: S100A2 is a calmodulin-like, p53-inducible, homodimeric protein that is readily oxidized in keratinocytes subjected to oxidative stress. Here we compare the redox status and subcellular distribution of S100A2 in normal human keratinocytes, immortalized keratinocytes (HaCaT), and malignant keratinocytes (A431) as a function of oxidative stress and intracellular Ca2+ levels. Normal human keratinocytes displayed strong nuclear and moderate cytoplasmic S100A2 immunoreactivity. HaCaT and A431 cells, which lack normal p53, expressed S100A2 in similar patterns but in 4- to 8-fold lower amounts. H2O2 treatment of normal human keratinocytes caused a reduction of nuclear S100A2 staining accompanied by an increase in cytoplasmic S100A2 staining, with a delayed time course (0.5-1 h) relative to S100A2 oxidative crosslinking (15 min). This phenomenon, consistent with translocation of S100A2 from the nucleus to the cytoplasm, could also be induced in normal human keratinocytes by increasing intracellular Ca2+ levels with the ionophore A23187. Sulfhydryl reducing agents blocked these changes, whether induced by H2O2 or increased intracellular Ca2+ levels. A temporal correlation was identified between S100A2 translocation at 1 h and loss of cell viability at 24 h after H2O2 treatment. A431 and HaCaT cells were strongly resistant to H2O2-induced S100A2 crosslinking, S100A2 translocation, and cell death. Increased intracellular Ca2+ levels caused prominent translocation of S100A2 in normal human keratinocytes and HaCaT, but not in A431 cells. These results identify S100A2 oxidation and translocation as markers for early cellular responses to oxidative stress, which are markedly attenuated in immortalized and malignant keratinocytes.
authors.affiliation: Department of Dermatology, University of Michigan, Ann Arbor 48109, USA.
authors.forename: Tong
authors.initials: T
authors.lastname: Zhang
authors.forename: Timothy L
authors.initials: TL
authors.lastname: Woods
authors.forename: James T
authors.initials: JT
authors.lastname: Elder
mesh_terms: D015415:Biomarkers; D002118:Calcium / Q000378:metabolism*; D002461:Cell Line, Transformed; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002630:Chemotactic Factors / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism*; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress / Q000502:physiology*; D009418:S100 Proteins / Q000378:metabolism*; D012867:Skin / Q000166:cytology / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D015415:Biomarkers; D002630:Chemotactic Factors; D016877:Oxidants; D009418:S100 Proteins; C098997:S100A2 protein, human; D016159:Tumor Suppressor Protein p53; D006861:Hydrogen Peroxide; D002118:Calcium