Expression of translationally controlled tumor protein in heat-stressed human dental pulp cells

Abstract

ObjectiveThe aim of this study was to investigate the effects of heat stress on cell viability, translationally controlled tumor protein (TCTP) expression, and the effects of recombinant TCTP on heat-stressed human dental pulp cells (HDPCs). MethodsHDPCs were isolated from human teeth and cultured at 37°C. For heat stress, HPDCs were incubated at 43°C for 45min. After heat stress, recombinant TCTP were added to HDPCs and cultured for various periods of time at 37°C. Heat-treated cells were then analyzed by DNA staining with Hoechst 33258, MTT, and caspase 3 activity assays. TCTP expression level was assessed by real-time PCR and western blot analysis. ResultsHeat-treated cells displayed lower cell density and nuclear morphology resembling apoptotic body. Heat stress significantly decreased cell viability and induced activity of caspase 3. The effect of recombinant TCTP on pulp cell death from heat stress varied depending on each subject and TCTP concentration. Heat stress up-regulated TCTP mRNA expression level. In contrast, TCTP protein level remained unchanged. Recombinant TCTP did not affect TCTP mRNA expression but down-regulated TCTP protein in heat-treated cells. ConclusionsHeat stress induces caspase 3 activation and up-regulates TCTP mRNA expression in HDPCs. TCTP did not play a key role on pulp cell recovery from heat stress.