title: Compensatory effects of hOGG1 for hMTH1 in oxidative DNA damage caused by hydrogen peroxide.
abstract: OBJECTIVE
  To investigate the potential compensatory effects of hOGG1 and hMTH1 in the repair of oxidative DNA damage.
  
  
  METHODS
  The hOGG1 and hMTH1 gene knockdown human embryonic pulmonary fibroblast cell lines were established by lentivirus-mediated RNA interference. The messenger RNA (mRNA) levels of hOGG1 and hM1TH1 were analyzed by the real-time polymerase chain reaction, and 8-hydroxy-2'-deoxyguanosine (8-oxo-dG) formation was analyzed in a high-performance liquid chromatography-electrochemical detection system.
  
  
  RESULTS
  The hOGG1 and hMTH1 knockdown cells were obtained through blasticidin selection. After transfection of hOGG1 and hMTH1 small interfering RNA, the expression levels of the mRNA of hOGG1 and hMTH1 genes were decreased by 97.2% and 96.2%, respectively. The cells then were exposed to 100 μmol/L of hydrogen peroxide (H2O2) for 12 h to induce oxidative DNA damage. After H2O2 exposure, hMTH1 mRNA levels were increased by 25% in hOGG1 gene knockdown cells, whereas hOGG1 mRNA levels were increased by 52% in hMTH1 gene knockdown cells. Following the treatment with H2O2, the 8-oxo-dG levels in the DNA of hOGG1 gene knockdown cells were 3.1-fold higher than those in untreated HFL cells, and 1.67-fold higher than those in H2O2-treated wild-type cells. The 8-oxo-dG levels in hMTH1 gene knockdown cells were 2.3-fold higher than those in untreated human embryonic pulmonary fibroblast cells, but did not differ significantly from those in H2O2-treated wild-type cells.
  
  
  CONCLUSIONS
  Our data suggested that hOGG1 could compensate for hMTH1 during oxidative DNA damage caused by H2O2, whereas hMTH1 could not compensate sufficiently for hOGG1 during the process.
authors.affiliation: Key Laboratory of Genetics & Molecular Medicine of Shenzhen, Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, China. Electronic address: keyke@szu.edu.cn.
authors.forename: Yuebin
authors.initials: Y
authors.lastname: Ke
authors.affiliation: Key Laboratory of Genetics & Molecular Medicine of Shenzhen, Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, China.
authors.forename: Ziquan
authors.initials: Z
authors.lastname: Lv
authors.affiliation: Key Laboratory of Genetics & Molecular Medicine of Shenzhen, Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, China.
authors.forename: Xifei
authors.initials: X
authors.lastname: Yang
authors.affiliation: Key Laboratory of Genetics & Molecular Medicine of Shenzhen, Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, China.
authors.forename: Jianqing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Genetics & Molecular Medicine of Shenzhen, Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Huang
authors.affiliation: Key Laboratory of Genetics & Molecular Medicine of Shenzhen, Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, China.
authors.forename: Shuang
authors.initials: S
authors.lastname: Wu
authors.affiliation: Department of Pharmacology, Campbell University School of Medicine, Buies Creek, NC 27546, USA; Department of Biomedical Sciences and Pathobiology, Virginia Polytechnic Institute and State University, and Virginia Tech-Wake Forest University School of Biomedical Engineering and Sciences, Blacksburg, VA 24061, USA.
authors.forename: Y Robert
authors.initials: YR
authors.lastname: Li
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D002460:Cell Line; D004249:DNA Damage; D045647:DNA Glycosylases / Q000235:genetics / Q000378:metabolism*; D045643:DNA Repair Enzymes / Q000235:genetics / Q000378:metabolism*; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D015971:Gene Expression Regulation, Enzymologic; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity*; D016086:Lentivirus / Q000235:genetics; D016877:Oxidants / Q000633:toxicity*; D018384:Oxidative Stress / Q000187:drug effects*; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D014162:Transfection
chemical_list: D016877:Oxidants; D012333:RNA, Messenger; D000080242:8-Hydroxy-2'-Deoxyguanosine; D006861:Hydrogen Peroxide; D010744:Phosphoric Monoester Hydrolases; D045647:DNA Glycosylases; C479058:oxoguanine glycosylase 1, human; C078207:8-oxodGTPase; D045643:DNA Repair Enzymes; D003849:Deoxyguanosine
keywords: 8-oxo-dG; Compensatory effects; RNAi; hMTH1; hOGG1