ADAM28 dramatically regulates the biological features of human gingival fibroblasts.

Abstract

This study was to explore the effects of a disintegrin and metalloproteinase 28 (ADAM28) on the proliferation, differentiation, and apoptosis of human gingival fibroblasts (HGFs) and probable mechanism. After ADAM28 antisense oligodeoxynucleotide (AS-ODN) and sense oligodeoxynucleotide (S-ODN) were transfected into HGFs by Lipofectamine 2000, respectively, the expression discrepancies of ADAM28 among various groups were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blotting. Methabenzthiazuron (MTT) and cell-cycle assays were used to test the HGFs proliferation activity. Annexin V fluorescein isothiocyanate (FITC)/propidium iodide (PI) and alkaline phosphatase (ALP) analysis were performed separately to measure apoptosis and the cytodifferentiation standard. Immunocytochemistry and Western-blotting were carried out to determine the influence of ADAM28 AS-ODN on HGFs expressing core binding factor α1 (Cbfα1), cementum protein 1 (CEMP1), osteopontin (OPN) and dentin matrix protein 1 (DMP1). The AS-ODN group displayed the lowest expression level in HGFs, meanwhile the ADAM28 S-ODN group showed the highest. Furthermore, blocking of ADAM28 could inhibit the proliferation of HGFs, enhance HGFs differentiation and induce apoptosis of HGFs. Whereas, overexpression of ADAM28 generated the opposite effects and inhibited apoptosis. ADAM28 AS-ODN was able to notably suppress the expressions of Cbfα1 and CEMP1, and ADAM28 had positive correlations with cbfα1 and CEMP1. These provided conspicuous evidence that ADAM28 may play a crucial role in root development as a potential regulator of growth, differentiation, and apoptosis of HGFs.