Marchese Cinzia, Visco Vincenzo, Aimati Laura, Cardinali Giorgia, Kovacs Daniela, Buttari Brigitta, Bellocci Marinella, Torrisi Maria Rosaria, Picardo Mauro
Dipartimento di Medicina Sperimentale e Patologia, Università di Roma 'La Sapienza'; Istituto Dermatologico San Gallicano, Roma, Italy.
Exp Dermatol. 2003 Aug;12(4):497-505. doi: 10.1034/j.1600-0625.2002.120419.x.
Keratinocytes play a key role in the pathogenesis of allergic contact dermatitis (ADC) induced by the sensitizing agent nickel. We analyzed here the effects of treatment with nickel and of the pretreatment with zinc on HaCaT cells and primary human keratinocytes. Cell counting, 5-bromo-2'-deoxyuridine incorporation assay and adenosine triphosphate (ATP) bioluminescence detection showed that treatment with NiSO4 induced DNA synthesis and cell proliferation and that pretreatment with ZnSO4 was able to abrogate this proliferative effect. This nickel-induced cell growth appeared enhanced when primary human keratinocytes were co-cultured with fibroblasts. Western blot analysis demonstrated that nickel ions induced up-modulation of the expression of the keratinocyte growth factor receptors (KGFR) without affecting the keratinocyte differentiation, whereas the protein levels of the epidermal growth factor receptor (EGFR) and of its ligand transforming growth factor-alpha (TGF-alpha) appeared unmodified by the treatment. Double immunofluorescence showed that the effect of nickel on DNA synthesis was mainly exerted on KGFR expressing cells, suggesting that KGFR up-modulation could be required for the nickel-induced cell proliferation. These results indicate that KGFR and its ligands may play a role in the mechanism of action of nickel ions and in the protective effect of zinc pretreatment.
角质形成细胞在致敏剂镍诱导的过敏性接触性皮炎（ADC）发病机制中起关键作用。我们在此分析了镍处理以及锌预处理对HaCaT细胞和原代人角质形成细胞的影响。细胞计数、5-溴-2'-脱氧尿苷掺入试验和三磷酸腺苷（ATP）生物发光检测表明，硫酸镍处理可诱导DNA合成和细胞增殖，而硫酸锌预处理能够消除这种增殖效应。当原代人角质形成细胞与成纤维细胞共培养时，这种镍诱导的细胞生长似乎增强。蛋白质印迹分析表明，镍离子可诱导角质形成细胞生长因子受体（KGFR）表达上调，而不影响角质形成细胞分化，而表皮生长因子受体（EGFR）及其配体转化生长因子-α（TGF-α）的蛋白质水平在处理后未发生改变。双重免疫荧光显示，镍对DNA合成的影响主要作用于表达KGFR的细胞，这表明KGFR上调可能是镍诱导细胞增殖所必需的。这些结果表明，KGFR及其配体可能在镍离子的作用机制以及锌预处理的保护作用中发挥作用。
