We have studied the effect of 20-hydroxyecdysone (20E) on cellular proliferation in IAL-PID2 cell line established from imaginal wing discs of Plodia interpunctella. Flow cytometry analysis demonstrated that 20E induced an arrest of cells in G2 phase. To determine whether this arrest was due to an effect of 20E on cyclin expression, we cloned two cDNA fragments, named PcycA and PcycB, encoding, respectively, Plodia cyclins A and B. Using PcycA and PcycB probes, we have demonstrated that 20E induced a sharp decrease in the levels of cyclin A and B expression. Studies of induction pattern of Plodia HR3 transcription factor by 20E revealed that its induction preceded the decrease of cyclins transcripts. An exposure of cells to 20E in the presence of juvenile hormone (JH) led to a change in the kinetic of PHR3 induction and prevented both the decline of cyclin A and B expression and the G2 arrest. This effect of JH provides an additional argument for the existence of a correlation between cyclin transcripts level and G2 arrest. For the first time in insects, these findings bring evidence that ecdysteroids regulate cellular proliferation by acting on cell cycle regulators as cyclins.