BmFoxO Gene Regulation of the Cell Cycle Induced by 20-Hydroxyecdysone in BmN-SWU1 Cells

SIMPLE SUMMARY: Ecdysteroid titer determines the state of the cell cycle in silkworm (Bombyx mori) metamorphosis. However, the mechanism of this process is unclear. In this study, we reported that 20-Hydroxyecdysone (20E) can promote BmFoxO (Bombyx mori Forkhead box protein O) gene expression and induce BmFoxO nuclear translocation in BmN-SWU1 cells. Overexpression of the BmFoxO gene affects cell cycle progression, which results in cell cycle arrest in the G0/G1 phase as well as inhibition of DNA replication. Further investigations showed that the effect of 20E was attenuated after BmFoxO gene knockdown. The findings of this study confirmed that BmFoxO is a key mediator in the cell cycle regulation pathway induced by 20E. This suggests a novel pathway for ecdysteroid-induced cell cycle regulation in the process of silkworm metamorphosis, and it is likely to be conserved between Lepidoptera insects. ABSTRACT: Ecdysteroid titer determines the state of the cell cycle in silkworm (Bombyx mori) metamorphosis. However, the mechanism of this process is unclear. In this study, we demonstrated that the BmFoxO gene participates in the regulation of the cell cycle induced by 20-Hydroxyecdysone (20E) in BmN-SWU1 cells. The 20E blocks the cell cycle in the G2/M phase through the ecdysone receptor (EcR) and inhibits DNA replication. The 20E can promote BmFoxO gene expression. Immunofluorescence and Western blot results indicated that 20E can induce BmFoxO nuclear translocation in BmN-SWU1 cells. Overexpression of the BmFoxO gene affects cell cycle progression, which results in cell cycle arrest in the G0/G1 phase as well as inhibition of DNA replication. Knockdown of the BmFoxO gene led to cell accumulation at the G2/M phase. The effect of 20E was attenuated after BmFoxO gene knockdown. These findings increase our understanding of the function of 20E in the regulation of the cell cycle in B. mori.