Transcriptional regulation of human amelotin gene by interleukin‐1β

One of the major causes of tooth loss is chronic inflammation of the periodontium, the tissues surrounding the tooth. Amelotin ( AMTN ) is a tooth enamel protein which is expressed in maturation‐stage ameloblasts and also in the internal basal lamina of junctional epithelium, a unique epithelial structure attached to the tooth surface which protects against the constant microbiological challenge to the periodontium. Localization of AMTN suggests that its function could be involved in the dentogingival attachment. The purpose of this study was to investigate the effect of interleukin‐1β ( IL ‐1β) on AMTN gene transcription in human gingival epithelial Ca9‐22 cells. IL ‐1β increased AMTN mRNA and protein levels at 3 h, and the levels reached maximum at 6 and 12 h. IL ‐1β induced luciferase activities of human AMTN gene promoter constructs (−211, −353, −501, −769, and −950AMTN), but these activities were partially inhibited in −353AMTN constructs that included 3‐bp mutations in CCAAT /enhancer binding protein 1 (C/ EBP 1), C/ EBP 2, and Ying Yang 1 ( YY 1) elements. Transcriptional activities induced by IL ‐1β were abrogated by protein kinase A ( PKA ), tyrosine kinase, mitogen‐activated protein kinase kinase ( MEK 1/2), and phosphatidylinositol 3‐kinase ( PI 3K) inhibitors. Gel shift and ChIP assays showed that IL ‐1β increased C/ EBP β binding to C/ EBP 1 and C/ EBP 2, and YY 1 binding to YY 1 elements after 3 h, and that these DNA –protein interactions were inhibited by PKA , tyrosine kinase, MEK 1/2, and PI 3K inhibitors. These results demonstrated that IL ‐1β increases AMTN gene transcription in human gingival epithelial cells mediated through C/ EBP 1, C/ EBP 2, and YY 1 elements in the human AMTN gene promoter.