title: [The role of ouabain in cell death of vascular endothelial cells ECV304 and the changes of expression of Na+, K(+)-ATPase alpha1, beta1-subunit].
abstract: OBJECTIVE
  To study the effect of Na+, K(+)-ATPase inhibition by ouabain on growth and death of vascular endothelial cells ECV304 and involved mechanisms.
  
  
  METHODS
  Growth inhibition of ouabain on ECV304 cells was analyzed using MTT assay. The feature of cell death was studied by Hoechst 33342/PI staining, transmission electron microscopy and DNA agarose gel electrophoresis in ECV304 cells treated with ouabain. The mRNA expression of Na+, K(+)-ATPase alpha1, beta1-subunit was examined by reverse transcription PCR (RT-PCR).
  
  
  RESULTS
  Ouabain inhibited the growth of ECV304 cells in a dose and time-dependent manner. 10 micromol/L ouabain treated for 24 hours could stimulate the necrosis of ECV304 cells; When treated with 0.1 micromol/L ouabain for 24-48 hours, the cells showed obviously defluxion, the loss of cell-cell contacts, nuclear chromatin condensation, chromatin margination and DNA fragmentation. Na+, K(+)-ATPase alpha1-subunit mRNA expression was significantly up-regulated in ECV304 cells treated with ouabain while the beta1-subunit expression conversely showed a significant decrease.
  
  
  CONCLUSIONS
  Ouabain could up-regulate Na+, K(+)-ATPase alpha1-Subunit expression and reduce beta1-Subunit expression which mediated signal transduction and decreased cell-cell adhesions and induced ECV304 cells death.
authors.affiliation: Department of Cellular Biology, Medical College of the Chinese People's Armed Police Forces, Tianjin 300162, China.
authors.forename: Na
authors.initials: N
authors.lastname: Wang
authors.forename: Rui-Cheng
authors.initials: RC
authors.lastname: Xu
authors.forename: Xiao-Yi
authors.initials: XY
authors.lastname: Chen
authors.forename: Wen-Liang
authors.initials: WL
authors.lastname: Hu
mesh_terms: D016923:Cell Death / Q000187:drug effects*; D002460:Cell Line; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D010042:Ouabain / Q000494:pharmacology*; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*
chemical_list: C503551:ATP1B1 protein, human; D010042:Ouabain; C499076:ATP1A1 protein, human; D000254:Sodium-Potassium-Exchanging ATPase