Highly synchronous culture of fibroblasts from G2 block caused by staurosporine, a potent inhibitor of protein kinases

The effect of staurosporine, a potent microbial inhibitor of protein kinases, on the cell cycle of cultured fibroblast cells was investigated. A low concentration of staurosporine (1–10 ng/ml) blocked the cell cycle of rat 3Y1 fibroblasts at the early G1 phase within 2 h after serum stimulation. On the other hand, a higher concentration of the drug (100 ng/ml) caused the specific G2 block. Both of these blocks were reversible. After release from the G2 block, highly synchronous transition to M phase was observed and both nuclear and cell divisions were completed within 180 min. This reversible G2 block showed a clear contrast to those by the other G2 arresters, trichostatin A and leptomycin B, which formed proliferative tetraploid cells after release by entering the cells into a new S phase without passage through M phase. The presence of trichostatin A or leptomycin B did not interfere with this synchronous progression through G2 M phases, suggesting that the arrest point of staurosporine was present in late G2 phase following those of trichostatin A and leptomycin B.