Tributyltin and dexamethasone induce apoptosis in rat thymocytes by mutually antagonistic mechanisms

The nuclei of apoptotic thymocytes can be identified by flow cytometry as a subpopulation exhibiting reduced DNA content. We observed that rat thymocyte cultures exposed to 1.0-2.5 microM tri-n-butyltin methoxide (TBT) exhibited a rapid time- and concentration-dependent induction of apoptosis, with > 85% of cells exhibiting reduced DNA content within 1 hr after exposure to 2.0-2.5 microM TBT. In contrast, exposure to 1.0 microM dexamethasone phosphate (DEX) resulted in a gradual time-dependent increase to approximately 45% induction of apoptosis by 6 hr versus approximately 15% spontaneous induction in controls. However, simultaneous exposure to TBT and DEX resulted in a decreased response: although TBT concentrations between 0.1 and 0.5 microM did not induce apoptosis, they reduced the ability of DEX to initiate apoptosis; while at TBT concentrations > or = 1.0 microM, simultaneous exposure to DEX substantially decreased the extent of TBT-induced apoptosis and cytotoxicity. Furthermore, while treatment with the protein synthesis inhibitor cycloheximide or the protein kinase C inhibitor H-7 completely blocked DEX-induced apoptosis, neither significantly reduced induction of apoptosis by TBT. Taken together, the toxicant-specific differences in the timing and extent of apoptotic induction and the dissimilar responses to CHX and H-7 suggest that TBT and DEX initiate endonuclease-mediated apoptotic cell death through different mechanisms. Moreover, the ability of each agent to retard the action of the other suggests that these mechanisms are directly or indirectly antagonistic.