title: Impact of JNK1, JNK2, and ligase Itch on reactive oxygen species formation and survival of prostate cancer cells treated with diallyl trisulfide.
abstract: OBJECTIVE
  In our previous study, we demonstrated that diallyl trisulfide (DATS) induced iron-dependent G2-M arrest of prostate cancer cell cycle. Moreover, ferritin degradation and an increase of labile iron pool has been linked to the activation of the JNK signaling axis. In the present work, we extended this study to determine which of the c-jun kinases is responsible for ferritin degradation and the role of iron in DATS-induced cell death. We hypothesized that JNK1 activates Itch ligase which will lead to ferritin ubiquitination, an increase in iron-dependent ROS formation and cell death.
  
  
  METHODS
  PC-3 prostate cancer cells were used in this study. Cell viability, concentration of ROS, labile iron pool, and changes in ferritin and P-Itch and DNA damage were determined.
  
  
  RESULTS
  We observed that DATS induced ferritin degradation through JNK, Itch signaling axis. DATS did not induce neither ROS formation nor increase the LIP in JNK1-DN transfected cells. We also observed that DATS increased JNK-dependent activating phosphorylation of E3ligase Itch. The cells transfected with inactive form of Itch were more resistant against cytotoxicity of DATS and showed lower DATS-induced ferritin degradation. Desferrioxamine a specific iron chelator had no effect neither on cell viability nor DNA damage evaluated by comet assay.
  
  
  CONCLUSIONS
  These results suggest that JNK1-dependent increase in LIP is mediated by Itch ubiquitin ligase.
authors.affiliation: Department of Bioenergetics and Physiology of Exercise, Medical University of Gdansk, Debinki 1, 80-211, Gdansk, Poland.
authors.forename: Alicja
authors.initials: A
authors.lastname: Sielicka-Dudzin
authors.forename: Andzelika
authors.initials: A
authors.lastname: Borkowska
authors.forename: Anna
authors.initials: A
authors.lastname: Herman-Antosiewicz
authors.forename: Michal
authors.initials: M
authors.lastname: Wozniak
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Jozwik
authors.forename: Donatella
authors.initials: D
authors.lastname: Fedeli
authors.forename: Jedrzej
authors.initials: J
authors.lastname: Antosiewicz
mesh_terms: D000498:Allyl Compounds / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D020552:Comet Assay; D004249:DNA Damage / Q000187:drug effects; D003676:Deferoxamine / Q000378:metabolism; D005293:Ferritins / Q000378:metabolism; D006801:Humans; D007501:Iron / Q000378:metabolism; D008297:Male; D048055:Mitogen-Activated Protein Kinase 8 / Q000235:genetics / Q000378:metabolism*; D048056:Mitogen-Activated Protein Kinase 9 / Q000235:genetics / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects; D010766:Phosphorylation; D011471:Prostatic Neoplasms / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013440:Sulfides / Q000494:pharmacology*; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*
chemical_list: D000498:Allyl Compounds; D017382:Reactive Oxygen Species; D012097:Repressor Proteins; D013440:Sulfides; C042577:diallyl trisulfide; D005293:Ferritins; D007501:Iron; C432966:ITCH protein, human; D044767:Ubiquitin-Protein Ligases; D048056:Mitogen-Activated Protein Kinase 9; D048055:Mitogen-Activated Protein Kinase 8; D003676:Deferoxamine