The regulation of Oct4 in human gingival fibroblasts stimulated by cyclosporine A: Preliminary observations.

BACKGROUND/PURPOSE: Oct4, a key transcription factor, could reprogram human somatic fibroblasts into embryonic stem cell-like pluripotent cells. The exact mechanism of cyclosporine A (CsA)-induced gingival overgrowth is still unclear. The aim of this study was to investigate the effects of CsA on the expression of Oct4 in cultured human gingival fibroblasts (HGFs) .

MATERIALS AND METHODS

The effects of CsA on HGFs were used to elucidate whether Oct4 expression could be induced by CsA by using quantitative real-time reverse transcription-polymerase chain reaction and western blot. Cell growth in CsA-treated HGFs with Oct4 lentiviral-mediated shRNAi knockdown was evaluated by tetrazolium bromide reduction assay.

RESULTS

CsA was found to upregulate Oct4 transcript in a dose-dependent manner (p < 0.05). CsA also dose-dependently increased Oct4 protein expression (p < 0.05). The lentivirus expressing sh-Oct4 successfully prevented the CsA-induced Oct4 mRNA and protein in HGFs (p < 0.05). However, knockdown of Oct4 was insufficient to inhibit CsA-stimulated cell growth in HGFs. Furthermore, double knockdown with pluripotency-associated transcription factor Nanog showed that the down-regulation of Oct4/Nanog by lentiviral infection significantly inhibited CsA-stimulated cell growth (p < 0.05).

CONCLUSION

Taken together, CsA was first found to upregulate Oct4 mRNA and protein expression in HGFs. The silencing Oct4 could not suppress cell growth unless Nanog was repressed simultaneously.