Regulation of calprotectin expression by interleukin‐1α and transforming growth factor‐β in human gingival keratinocytes
2007
Background and Objective: Calprotectin, a heterodimer of S100A8 and S100A9 with antimicrobial properties, is expressed in gingival keratinocytes and plays an important role in innate immunity. Because calprotectin expression is localized in the spinous cell layer of the gingival epithelium, we hypothesized that the expression of calprotectin in keratinocytes is related to the differentiation stage. The aim of the present study was to investigate the relationship between calprotectin expression and keratinocyte differentiation using some factors that regulated its differentiation.
Material and Methods: Normal human gingival keratinocytes were isolated from gingival tissues obtained at the extraction of wisdom teeth, and were cultured in serum-free keratinocyte medium supplemented with interleukin-1α or calcium, which promote keratinocyte differentiation, and transforming frowth factor-β (TGF-β) or retinoic acid, which suppress its differentiation. The expression of S100A8/A9 mRNA and the production of calprotectin in normal human gingival keratinocytes were examined by northern blotting and enzyme-linked immunosorbent assay, respectively. The expression of cytokeratin 14, involucrin and filaggrin (marker proteins of keratinocyte differentiation) was investigated by immunohistochemical staining, and the DNA-binding activity of CCAAT/enhancer binding protein α (C/EBPα), a transcription factor, was examined by electrophoretic mobility shift assay.
Results: The expression of S100A8/A9 mRNA and the production of calprotectin were increased by interleukin-1α and calcium, but decreased by TGF-β. RA inhibited the expression of S100A8/A9 and keratinocyte differentiation, which were induced by interleukin-1α. C/EBPα DNA-binding activity in normal human gingival keratinocytes was enhanced by interleukin-1α and calcium, but suppressed by TGF-β.
Conclusion: The present study suggests that calprotectin expression is related to keratinocyte differentiation and that C/EBPα is a regulator of calprotectin expression in keratinocytes.
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