Cooperative function of Chk1 and p38 pathways in activating G2 arrest following exposure to temozolomide

Object. The Chk1 and p38 mitogen-activated protein kinase (MAPK) pathways play key roles in the G(2) arrest caused by exposing glioma cells to temozolomide (TMZ). Although inhibition of either pathway sensitizes glioma cells to TNZ-induced cytotoxicity, the relative contributions of these pathways to TMZ-induced G(2) arrest and to TMZ resistance conferred by G(2) arrest have not been defined. Methods. The authors pharmacologically inhibited the Chk1 and/or p38 pathways in U87MG human glioma cells prior to and/or after exposure to TMZ; thereafter, effects on the TNZ-induced G(2) arrest pathway and toxicity were monitored. The p38 inhibitor SB203580 or the Chk1 inhibitor UCN-01 or their combination blocked TMZ-mediated inactivation of cdc25C and cdc2, suggesting that p38 and Chk1 pathways work cooperatively and are both necessary to inactivate cdc25C and cdc2. Consistent with this idea, the inhibition of both Chk1 and p38 pathways did not lead to greater bypass of TMZ-induced G(2) arrest or greater cytotoxicity than inhibition of either pathway alone. Inhibition of p38 did not alter TMZ-induced Chk1 activation/phosphorylation and vice versa, suggesting that p38 and Chk1 do not cooperatively bring about G(2) arrest by reciprocal activation/phosphorylation. The two pathways, however, are not functionally identical; the Chk1 pathway was required for both the initiation and maintenance of TMZ-induced G(2) arrest, whereas the p38 pathway played a role only in the initiation. Conclusions. The Chk1 and p38 pathways cooperate to bring about TMZ-induced G(2) arrest, and the inhibition of either pathway alone is sufficient to sensitize U87MG glioma cells to TMZ-induced cytotoxicity.