Objective To explore the effect of simulated hypoxic environment on proliferation, cell cycle, apoptosis and the protein and mRNA expressions of SOX2 and OCT4 in tongue squamous cell carcinoma SCC-15 cells. Methods SCC-15 cells was cultured in vitro, and the hypoxia mimetic agent desferrioxamine (DFO) was added to simulate hypoxic condition (simulated hypoxia group, HOX group) and normoxia was designed as control group (NOX group). CCK-8 assay was performed to measure the proliferation ability of the SCC-15 cells; flow cytometry was used to measure the apoptosis rate and cell cycle of the SCC-15 cells; Western blotting was applied to measure the expressions of HIF-1α, SOX2 and OCT4 protein in NOX and HOX groups; qPCR was used to measure the expressions of HIF-1α, SOX2 and OCT4 mRNA in NOX and HOX groups. Results DFO effectively simulated the hypoxic condition. Compared with the NOX group, the proliferation ability of the cells in HOX group was significantly inhibited by DFO (P<0.05), the proportion in G₁ cells was increased and that in S+G₂ was decreased; while no significant difference was found in the apoptosis rate of SCC-15 cells; the expressions of HIF-1α, SOX2 and OCT4 proteins were significantly increased in HOX group, and the expressions of HIF-1α and OCT4 proteins were significantly higher than SOX2 (P<0.05). There was no significant difference of mRNA expression of HIF-1α in NOX and HOX groups (P>0.05); the mRNA expressions of SOX2 and OCT4 in HOX group were significantly higher than those in NOX group (P<0.05). Conclusion DFO can effectively simulate hypoxic environment, and hypoxia can promote the expressions of SOX2 and OCT4 proteins and mRNA.