title: [Study on the effect of cell proliferation and anti-oxidative damage of aldehyde dehydrogenase-2 gene transfected into K562 cells].
abstract: OBJECTIVE
  To construct a eukaryotic expression vector containing aldehyde dehydrogenase-2 (ALDH2) gene and investigate the effects and its possible mechanisms of ALDH2 gene on cell proliferation and anti-oxidative damage in the K562 cells.
  
  
  METHODS
  An eukaryotic expression vector containing the ALDH2 gene cloned from human hepatocytes was constructed and transfected into K562 cells by liposome. RT-PCR and Western blot were used to evaluate the expression of ALDH2. MTT assay was used to check the cell proliferation and trypan blue exclusion to check K562 cells damage induced by hydrogen peroxide (H2O2). RT-PCR and fluorescence spectrophotometry were used to determine the expression of heme oxygenase-1 (HO-1) and the generation of intracellular reactive oxygen species (ROS) respectively.
  
  
  RESULTS
  RT-PCR and Western blot analysis showed distinct higher ALDH2 protein expression in gene transfected group. The latter group had a higher cell proliferation (P < 0.05) and survival rate against H2O2 induced-oxidative damage, being increased by 7.8 times (IC(50) was 12.3 µmol/L and 1.4 µmol/L for K562-pcDNA3.1-ALDH2 and control cells, respectively, P < 0.01). The HO-1 mRNA expression and the generation of intracellular ROS were downregulated at a specific concentration of H2O2 in the ALDH2 gene transfected group.
  
  
  CONCLUSIONS
  ALDH2 gene transfection can protect K562 cells against oxidative damage, and the downregulation of HO-1 expression and intracellular ROS may be involved in this process.
authors.affiliation: Department of Hematology, The Affiliated Hospital of Guiyang Medical College, Guiyang 550004, China.
authors.forename: Ji-Shi
authors.initials: JS
authors.lastname: Wang
authors.forename: Xiu-Ying
authors.initials: XY
authors.lastname: Hu
authors.forename: Qin
authors.initials: Q
authors.lastname: Fang
authors.forename: Jian-Qiong
authors.initials: JQ
authors.lastname: Xie
authors.forename: Yuan
authors.initials: Y
authors.lastname: Yang
authors.forename: Xin
authors.initials: X
authors.lastname: Cui
authors.forename: Bai-Sheng
authors.initials: BS
authors.lastname: Chai
mesh_terms: D000444:Aldehyde Dehydrogenase; D017209:Apoptosis / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D006801:Humans; D006861:Hydrogen Peroxide; D020014:K562 Cells; D012333:RNA, Messenger / Q000235:genetics; D014162:Transfection
chemical_list: D012333:RNA, Messenger; D006861:Hydrogen Peroxide; D000444:Aldehyde Dehydrogenase