Hydrogen sulfide inhibits cell proliferation and induces cell cycle arrest via an elevated p21Cip1 level in Ca9-22 cells

Background and Objective: Volatile sulfur compounds such as hydrogen sulfide (H2S) and methyl mercaptan (CH3SH) are the main causes of oral malodor. However, the physiological functions of H2S have not been investigated in oral tissues. The aim of this study was to evaluate the effect of H2S on cell proliferation and the cell cycle in oral epithelial-like cells. Material and Methods: Ca9-22 cells were used in this study. Cells were cultured in 5% CO2/95% air with (5 or 10 ng/mL) or without H2S. DNA synthesis was measured using a 5-bromo-2-deoxyuridine enzyme-linked immunosorbent assay. The cell cycle was analyzed using a flow cytometer. The expressions of phosphorylated retinoblastoma protein (Rb), p21(Cip1) and p27(Kip1) were evaluated by western blotting. Results: Exposure to 5 and 10 ng/mL of H2S significantly decreased DNA synthesis (p < 0.05). Cell cycle analysis also showed that exposure to both concentrations of H2S significantly increased the proportion of cells in G(1) phase (p < 0.001) and significantly decreased the proportion of cells in S phase (p < 0.01). Western blotting showed that Rb phosphorylation was reduced and p21(Cip1) was enhanced by exposure to H2S. Conclusion: The results indicated that H2S inhibits cell proliferation and induces cell cycle arrest via the expression of p21(Cip1) in Ca9-22 cells.