Abstract
Purpose
We evaluated in vitro the role of CO2-induced oxidative stress on the expression of proteins involved in cell-cycle regulation of neuroblastoma cells.
Methods
SH-SY5Y cells were exposed to CO2 at 15 mmHg pressure (100 %) for 4 h and then moved to normal condition for 24 h. Control cells were maintained in 5 % CO2 for the same time. ROS production was determined by fluorescent staining with H2DCF-DA. DNA damage was measured by COMET assay. p53 protein expression was analyzed by western blot and confocal laser scanning microscopy was used to evaluate its sub-cellular localization. Cyclin expression was quantified by real-time PCR and western blot. Cell-cycle analysis was performed by FACS.
Results
CO2 incubation was associated with an increase in ROS production (p < 0.01), cell DNA damage mainly after 24 h (12 % increase of tail DNA content and 4-fold increase of tail length) and a significant up-regulation in p53 expression at 24 h with an intense nuclear staining. In CO2-treated cells, we observed an S-phase arrest in correlation with a reduction of cyclin B1 expression.
Conclusions
In vitro-simulated pneumoperitoneum environment with CO2 induces oxidative stress and cell DNA damage, leading to p53 up-regulation involved in cell-cycle arrest of neuroblastoma cells.
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Montalto, A.S., Currò, M., Russo, T. et al. In vitro CO2-induced ROS production impairs cell cycle in SH-SY5Y neuroblastoma cells. Pediatr Surg Int 29, 51–59 (2013). https://doi.org/10.1007/s00383-012-3206-3
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DOI: https://doi.org/10.1007/s00383-012-3206-3