title: Neutrophil gelatinase-associated lipocalin acts as a protective factor against H(2)O(2) toxicity.
abstract: BACKGROUND
  Lipocalin 2 (Lcn2, NGAL) is a member of the lipocalin superfamily for which a variety of functions have been reported. However, the precise biological roles of NGAL are not fully known. We have investigated the ability of NGAL to prevent H(2)O(2) toxicity, which is considered to be the classical inducer of oxidative stress caused by ROS generation in an in vitro model.
  
  
  METHODS
  NGAL cDNA was isolated from HepG2 cell line and cloned to pcDNA3.1(+) vector. The construct was transfected to CHO cell line. Stable clones were generated, and the expression of NGAL was determined by RT-PCR, Western blot analysis and ELISA. NGAL gene in A549 cell line was downregulated with the siRNA. CHO and A549 cells were intoxicated with H(2)O(2) and cell proliferation was performed by MTT assay. Apoptotic cells were detected by flow cytometry.
  
  
  RESULTS
  Cell proliferation was higher in CHO expressing NGAL in doses of 5 and 10 mM H(2)O(2) after 2h compared with the control. H(2)O(2) was also more toxic in the presence of NGAL siRNA compared with the control in A549 cell. Our results also revealed that NGAL protect cells from apoptosis.
  
  
  CONCLUSIONS
  Overall, our results revealed for the first time a new function for NGAL/Lcn2: acting as a protective factor against H(2)O(2) toxicity. In the future, NGAL may have the potential application to ameliorate the toxicity induced by oxidative stress conditions.
authors.affiliation: Research Center, Iranian Blood Transfusion Organization, Tehran Iran. Roudkenar@ibto.ir
authors.forename: Mehryar Habibi
authors.initials: MH
authors.lastname: Roudkenar
authors.forename: Raheleh
authors.initials: R
authors.lastname: Halabian
authors.forename: Zahra
authors.initials: Z
authors.lastname: Ghasemipour
authors.forename: Amaneh Mohammadi
authors.initials: AM
authors.lastname: Roushandeh
authors.forename: Mahdi
authors.initials: M
authors.lastname: Rouhbakhsh
authors.forename: Mahin
authors.initials: M
authors.lastname: Nekogoftar
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Kuwahara
authors.forename: Manabu
authors.initials: M
authors.lastname: Fukumoto
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Shokrgozar
mesh_terms: D000209:Acute-Phase Proteins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis; D001483:Base Sequence; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D017931:DNA Primers; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D000071068:Lipocalin-2; D054834:Lipocalins / Q000235:genetics / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering
chemical_list: D000209:Acute-Phase Proteins; D017931:DNA Primers; C489506:LCN2 protein, human; D000071068:Lipocalin-2; D054834:Lipocalins; D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; D006861:Hydrogen Peroxide