title: [Effect of AKR1A1 knock-down on H2;O2; and 4-hydroxynonenal-induced cytotoxicity in human 1321N1 astrocytoma cells].
abstract: OBJECTIVE
  To investigate the role of human aldo-keto reductase 1A1 (AKR1A1) in the resistance to oxidative stress and the metabolism of toxic aldehyde in astrocytoma cells.
  
  
  METHODS
  The siRNA was transfected into 1321N1 astrocytoma cells using Lipofectamine(TM); RNAiMax. Western blotting and qRT-PCR were applied to evaluate the knock-down efficiency of AKR1A1. MTT assay was used to examine the cell viability after H2;O2; and 4-hydroxynonenal treatment in AKR1A1 knock-down cells. In addition, the effect of knocking down AKR1A1 on cellular reactive oxygen species (ROS) level in the presence of H2;O2; was measured using 2', 7'-dichlorofluorescein (DCFH-DA).
  
  
  RESULTS
  Western blotting and qRT-PCR showed that the AKR1A1-specific siRNA inhibited AKR1A1 gene expression by about 70% in 1321N1 cells. Cells with knock-down of AKR1A1 were more sensitive to H2;O2; and 4-hydroxynonenal-induced cytotoxicity. Furthermore, cellular ROS level in the cells with knock-down of AKR1A1 was much higher than that in the control cells in the presence of H2;O2;.
  
  
  CONCLUSIONS
  The specific siRNA could efficiently inhibit AKR1A1 expression in 1321N1 cells. AKR1A1 could be involved in the metabolism of 4-hydroxynonenal and play a role in the resistance to oxidative stress.
authors: School of Pharmacy, China Medical University, Shenyang, China. danlicmu@hotmail.com
authors: Dan
authors: D
authors: Li
authors: Qishan
authors: Q
authors: Zhang
authors: Lina
authors: L
authors: Zhou
authors: Rong
authors: R
authors: Liu
mesh_terms: D000449:Aldehyde Reductase / Q000172:deficiency* / Q000235:genetics* / Q000378:metabolism; D000447:Aldehydes / Q000633:toxicity*; D001254:Astrocytoma / Q000473:pathology*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics; D055785:Gene Knockdown Techniques; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism / Q000633:toxicity*; D018384:Oxidative Stress / Q000187:drug effects / Q000235:genetics; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D000447:Aldehydes; D012333:RNA, Messenger; D006861:Hydrogen Peroxide; C580929:AKR1A1 protein, human; D000449:Aldehyde Reductase; C027576:4-hydroxy-2-nonenal