Inhibition of G1 cell cycle arrest in human gingival fibroblasts exposed to phenytoin

Gingival overgrowth is caused in response to the antiepileptic drug phenytoin (PHT). PHT-induced gingival overgrowth is characterized by the proliferation of fibroblasts and increased collagen formation in gingiva. Fibroblast proliferation is regulated through the cell cycle. Thus, in the present study, we examined the effects of PHT on the cell cycle, the expression of cell cycle control proteins and the proliferation in human gingival fibroblasts (hGFs). Cells were stimulated in serum-free DMEM with or without 0.25 mu m PHT. Subsequently, the cell cycle phase distribution and the protein expression after 24h and the cell proliferation after 24, 48 and 72h were evaluated. PHT significantly inhibited synchronization at the G(0)/G(1) phase of the cell cycle in hGFs through serum starvation. Stimulation with PHT for 48 and 72h significantly induced a proliferative response in hGFs. PHT decreased the expression of the Cdk-inhibitory proteins p21 and p27 and increased the levels of the S phase-promoting proteins phospho-Thr160-Cdk2 and phospho-Ser807/811-Rb in serum-free DMEM. The inhibition of G(1) cell cycle arrest in hGFs may result from an increase in phosphorylated Cdk2 and Rb proteins and decreased levels of p21 and p27 proteins by PHT. The gingival overgrowth may be caused by the failure of the G1 cell cycle arrest in GFs exposed to PHT.