To understand antitumor activity of an edible mushroom Paecilomyces japonica, we have investigated its effect on the cell cycle. When Jurkat T cells transfected with vector (JT/Neo) or Bcl-2 gene (JT/Bcl-2) were treated with the ethyl acetate extract (4–6μg/ml) of P. japonica for 40h, JT/Neo cells underwent apoptosis with no detectable G 1 -arrest, whereas JT/Bcl-2 cells that failed to induce apoptosis accumulated at the G 1 along with reduction of hyperphosphorylated Rb. The cdk4, cyclin E and A, required for the G 1 -cdks that phosphorylate Rb, were markedly downregulated in JT/Bcl-2 cells, and the G 1 -cdk inhibitor p27 Kip1 was significantly upregulated. GC–MS analysis identified a component structurally similar to diacetoxyscirpenol as the effective ingredient contributing to the G 1 -arrest. These results demonstrate that P. japonica can arrest the cell cycle of JT/Bcl-2 cells at the G 1 by suppressing phosphorylation of Rb through downregulation of the activity of G 1 -cdks, and thus potentiates apoptotic cell death.