In vitro CO sub(2) -induced ROS production impairs cell cycle in SH-SY5Y neuroblastoma cells

• Published in: Pediatric surgery international 2013-01, Vol.29 (1), p.51-59
• Main Authors: Montalto, Angela Simona, Curro, Monica, Russo, Tiziana, Visalli, Giuseppa, Impellizzeri, Pietro, Antonuccio, Pietro, Arena, Salvatore, Borruto, Francesca Astra, Scalfari, Gianfranco, Ientile, Riccardo, Romeo, Carmelo
• Format: Article
• Language: English
• ISSN: 0179-0358; 1437-9813
• DOI: 10.1007/s00383-012-3206-3

Purpose: We evaluated in vitro the role of CO sub(2) -induced oxidative stress on the expression of proteins involved in cell-cycle regulation of neuroblastoma cells. Methods: SH-SY5Y cells were exposed to CO sub(2) at 15 mmHg pressure (100 %) for 4 h and then moved to normal condition for 24 h. Control cells were maintained in 5 % CO sub(2) 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: CO sub(2) 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 CO sub(2) -treated cells, we observed an S-phase arrest in correlation with a reduction of cyclin B1 expression. Conclusions: In vitro-simulated pneumoperitoneum environment with CO sub(2) induces oxidative stress and cell DNA damage, leading to p53 up-regulation involved in cell-cycle arrest of neuroblastoma cells.