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Animal studies have shown that irregular light-dark cycles cause circadian desynchronization, while few studies have addressed the effect of regular/irregular stimulation cycles of signaling hormones on the cellular clock  in vitro . Here, we examined how cellular clocks respond to regular and irregular stimulation cycles of dexamethasone, using NIH3T3 cells transfected with the  Bmal1  promoter-driven luciferase ( Bmal1-Luc ) reporter gene. Cyclic stimulation with dexamethasone at different time intervals (18–28 h, 3 times regularly) revealed that  Bmal1-Luc  bioluminescence rhythms can be entrained to 22 and 24 h cycles during the stimulation period, but not to other cycles. The rhythm entrained for 24 h cycles persisted for at least one day after the last stimulation. Irregular dexamethasone treatment (16, 24, and 16 h, sequentially; short-term jet lag protocol) resulted in an overall upregulation and phase shifts of the temporal expression of several clock genes and cell cycle genes, including  c-Myc  and  p53 . Regular dexamethasone stimulation three times with 24 h cycles also caused upregulation of  Per1  and  Per2  expression, but not  c-Myc  and  p53  expression. In conclusion, our study identified the entrainable range of the circadian clock in NIH3T3 cells to the dexamethasone stimulation cycle and demonstrated that irregular dexamethasone treatment could disturb the expression of cell cycle genes.

Effect of regular and irregular stimulation cycles of dexamethasone on circadian clock in NIH3T3 cells