title: Nitric oxide donating nonsteroidal anti-inflammatory drugs induce apoptosis in human prostate cancer cell systems and human prostatic stroma via caspase-3.
abstract: OBJECTIVE
  New nitric oxide (NO) donating nonsteroidal anti-inflammatory drugs (NSAIDs) have been synthesized to counteract the side effects of conventional NSAIDs. Mounting evidence suggests that NSAIDs may have a possible chemopreventative/therapeutic role in prostate cancer. NO is a powerful biological messenger with multiple cellular effects. We established the effects of 2 of these new drugs in prostate cell systems.
  
  
  METHODS
  We studied the effects of NO-ibuprofen (NCX 2111) and NO-aspirin (NCX 4060) on hormone sensitive (LNCap) and insensitive (PC3) prostate cancer epithelial cell lines as well as primary cultures of prostatic stroma. Proliferation was measured using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazalium bromide) assay to examine proliferation. Subsequently flow cytometry, cell cycle analysis and TUNEL staining were used to look for apoptosis. Caspase-3 expression was also examined in treated cell types.
  
  
  RESULTS
  NCX 2111 and NCX 4060 were found to be potent inhibitors of proliferation in a dose dependent fashion. The 2 drugs induced apoptosis, as seen by flow cytometry, cell cycle analysis and TUNEL staining, at doses between 10 and 100 microM. These NO-NSAIDs increased caspase-3 expression. NCX 4060 was more effective at lower concentrations (10 microM) but each compound was much more potent than conventional ibuprofen and aspirin at inducing apoptosis and inhibiting proliferation.
  
  
  CONCLUSIONS
  NO-NSAIDs are potent antiproliferative pro-apoptotic compounds in prostate cell systems. This pro-apoptotic effect is mediated via caspase-3 and it is independent of the type of prostate cell used. These findings have ramifications for the use of these new drugs in prostate cancer chemoprevention or treatment.
authors.affiliation: Department of Urology, Western General Hospital, Prostate Research Group, University of Edinburgh, Western General Hospital, Edinburgh, United Kingdom. justine.royle@virgin.net
authors.forename: Justine Sarah
authors.initials: JS
authors.lastname: Royle
authors.forename: James A
authors.initials: JA
authors.lastname: Ross
authors.forename: Ian
authors.initials: I
authors.lastname: Ansell
authors.forename: Prasad
authors.initials: P
authors.lastname: Bollina
authors.forename: David N
authors.initials: DN
authors.lastname: Tulloch
authors.forename: Fouad K
authors.initials: FK
authors.lastname: Habib
mesh_terms: D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001241:Aspirin / Q000031:analogs & derivatives* / Q000494:pharmacology*; D053148:Caspase 3; D020169:Caspases / Q000502:physiology*; D002455:Cell Division / Q000187:drug effects; D004396:Coloring Agents; D004305:Dose-Response Relationship, Drug; D044950:Flavanones / Q000138:chemical synthesis / Q000633:toxicity; D005434:Flow Cytometry; D006801:Humans; D007052:Ibuprofen / Q000031:analogs & derivatives* / Q000494:pharmacology*; D020287:In Situ Nick-End Labeling; D008297:Male; D009566:Nitrates / Q000494:pharmacology*; D020030:Nitric Oxide Donors / Q000494:pharmacology*; D011471:Prostatic Neoplasms / Q000188:drug therapy*; D013778:Tetrazolium Salts; D013844:Thiazoles; D014407:Tumor Cells, Cultured
chemical_list: C488175:2-(acetyloxy)benzoic acid 6-(nitrooxymethyl)-2-phenylmethyl ester; C462031:6-(1,1-dimethylallyl)naringenin; D000894:Anti-Inflammatory Agents, Non-Steroidal; D004396:Coloring Agents; D044950:Flavanones; C488176:N-acetyl-S-(alpha-methyl-4-(2-methylpropyl)benzeneacetyl)cysteine 4-(nitrooxy)butyl ester; D009566:Nitrates; D020030:Nitric Oxide Donors; D013778:Tetrazolium Salts; D013844:Thiazoles; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; C022616:thiazolyl blue; D001241:Aspirin; D007052:Ibuprofen