title: In vitro CO2-induced ROS production impairs cell cycle in SH-SY5Y neuroblastoma cells.
abstract: OBJECTIVE
  We evaluated in vitro the role of CO(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(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(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(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(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(2) induces oxidative stress and cell DNA damage, leading to p53 up-regulation involved in cell-cycle arrest of neuroblastoma cells.
authors.affiliation: Unit of Pediatric Surgery, Department of Pediatric, Gynecological, Microbiological and Biomedical Sciences, University of Messina, Policlinico G. Martino, Viale Gazzi, 98124, Messina, Italy.
authors.forename: Angela Simona
authors.initials: AS
authors.lastname: Montalto
authors.forename: Monica
authors.initials: M
authors.lastname: Currò
authors.forename: Tiziana
authors.initials: T
authors.lastname: Russo
authors.forename: Giuseppa
authors.initials: G
authors.lastname: Visalli
authors.forename: Pietro
authors.initials: P
authors.lastname: Impellizzeri
authors.forename: Pietro
authors.initials: P
authors.lastname: Antonuccio
authors.forename: Salvatore
authors.initials: S
authors.lastname: Arena
authors.forename: Francesca Astra
authors.initials: FA
authors.lastname: Borruto
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Scalfari
authors.forename: Riccardo
authors.initials: R
authors.lastname: Ientile
authors.forename: Carmelo
authors.initials: C
authors.lastname: Romeo
mesh_terms: D002245:Carbon Dioxide / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects*; D004249:DNA Damage / Q000187:drug effects; D006801:Humans; D009447:Neuroblastoma / Q000473:pathology*; D018384:Oxidative Stress / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D017382:Reactive Oxygen Species; D002245:Carbon Dioxide