title: JunD mediates androgen-induced oxidative stress in androgen dependent LNCaP human prostate cancer cells.
abstract: BACKGROUND
  Numerous and compelling evidence shows that high level of reactive oxygen species (ROS) plays a key role in prostate cancer occurrence, recurrence and progression. The molecular mechanism of ROS overproduction in the prostate gland, however, remains mostly unknown. Unique AP-1 transcription factor JunD has been shown to inhibit cell proliferation, promote differentiation and mediate stress responses in a variety of eukaryotic cells. We previously reported that androgen-androgen receptor induced ROS production in androgen-dependent LNCaP human prostate cancer cells is associated with increased JunD level/AP-1 transcriptional activity.
  
  
  METHODS
  LNCaP cells constitutively overexpressing a functionally inactive form of JunD (JunDDeltaTA) or stably transfected with JunD siRNA (siJunD) to suppress JunD protein expression were established. Overexpression of JunD in LNCaP cells using transient transfection method was applied to assess the induction of ROS production in LNCaP cells. DCF assay was used to measure the ROS concentrations in the transfected as well as non-transfected control cells. RT-PCR and Western blot analyses were used to confirm silencing or overexpression of JunD in the transfected cells.
  
  
  RESULTS
  In the absence of androgen, LNCaP cells transiently transfected with a JunD overexpressing vector have relatively enhanced cellular ROS levels as compared to LNCaP cells transfected with a vector control. LNCaP cells that fail to express functional JunD (JunDDeltaTA or siJunD) do not exhibit any increase in ROS production in response to androgen.
  
  
  CONCLUSIONS
  Based on these data, we conclude that JunD is an essential mediator of the androgen-induced increase in ROS levels in LNCaP cells.
authors.affiliation: University of Wisconsin Paul P. Carbone Comprehensive Cancer Center, University of Wisconsin, Madison, USA.
authors.forename: Farideh
authors.initials: F
authors.lastname: Mehraein-Ghomi
authors.forename: Elyse
authors.initials: E
authors.lastname: Lee
authors.forename: Dawn R
authors.initials: DR
authors.lastname: Church
authors.forename: Todd A
authors.initials: TA
authors.lastname: Thompson
authors.forename: Hirak S
authors.initials: HS
authors.lastname: Basu
authors.forename: George
authors.initials: G
authors.lastname: Wilding
mesh_terms: D015153:Blotting, Western; D045744:Cell Line, Tumor; D005822:Genetic Vectors; D006727:Hormone Antagonists / Q000494:pharmacology; D006801:Humans; D008297:Male; D015741:Metribolone / Q000494:pharmacology; D009376:Neoplasms, Hormone-Dependent / Q000235:genetics / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects / Q000235:genetics; D011471:Prostatic Neoplasms / Q000235:genetics / Q000378:metabolism*; D016755:Proto-Oncogene Proteins c-jun / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D012334:RNA, Neoplasm / Q000737:chemistry / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018808:Transcription Factor AP-1 / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic; D014162:Transfection
chemical_list: D006727:Hormone Antagonists; D016755:Proto-Oncogene Proteins c-jun; D012334:RNA, Neoplasm; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D018808:Transcription Factor AP-1; D015741:Metribolone