title: [Mechanisms for inhibitory effect of ALDH2 on doxorubicin-induced cytotoxicity in C2C12 myogenic cell line].
abstract: OBJECTIVE
  To investigate the mechanisms for inhibitory effect of aldehyde dehydrogenase 2 (ALDH2) on doxorubicin (DOX)-induced cytotoxicity in C2C12 myogenic cell line.
  
  
  METHODS
  Cell apoptosis was evaluated by flow cytometry and the activity of capase-3/7. The relative content of reactive oxygen species (ROS) and 4-hydroxynonenal (4-HNE) were detected by chemical fluorometric enzyme immunoassay. The protein and mRNA expression of ALDH2, Bcl-2, NADPH oxidase 2 (NOX2) and the cytoplasmic subunit p-p47PHOX were evaluated by Western blot and quantitative PCR, respectively.
  
  
  RESULTS
  Overexpression of ALDH2 attenuated DOX-induced cell toxicity (increase in apoptosis and inhibition of proliferation), which were reversed by downregulation of ALDH2. Overexpression of ALDH2 reduced p47PHOX phosphorylation levels, and suppressed activation of NOX2 and ROS production, which were reversed by downregulation of ALDH2. Moreover, apocynin, an inhibitor of NOX, reduced the cytotoxicity of DOX concomitantly with a decrease in phosphorylation of p47PHOX, ROS production and caspase-3/7 activity, and an increase in the activity and expression of ALDH2.
  
  
  CONCLUSIONS
  DOX-induced cytotoxicity is related to increase of intracellular oxidative stress, which is involved in unregulation of NOX2 and downregulation of ALDH2. Activation of ALDH2 could exert cytoprotection via inhibiting NOX2-dependent ROS production.
authors.affiliation: Department of Oncology, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Yawen
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Department of Geriatrics, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Shicong
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Department of Geriatrics, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Oncology, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Yufang
authors.initials: Y
authors.lastname: Li
authors.affiliation: Departments of Vasculocardiology,  Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Shenghua
authors.initials: S
authors.lastname: Zhou
authors.affiliation: Department of Oncology, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Xianling
authors.initials: X
authors.lastname: Liu
mesh_terms: D000071396:Aldehyde Dehydrogenase, Mitochondrial; D000447:Aldehydes; D000818:Animals; D017209:Apoptosis; D053148:Caspase 3; D002460:Cell Line; D002470:Cell Survival; D015536:Down-Regulation; D004317:Doxorubicin; D051379:Mice; D018384:Oxidative Stress; D010766:Phosphorylation; D017382:Reactive Oxygen Species
chemical_list: D000447:Aldehydes; D017382:Reactive Oxygen Species; D004317:Doxorubicin; C515029:ALDH2 protein, mouse; D000071396:Aldehyde Dehydrogenase, Mitochondrial; C505409:Casp3 protein, mouse; D053148:Caspase 3; C027576:4-hydroxy-2-nonenal