Nitric oxide donating nonsteroidal anti-inflammatory drugs induce apoptosis in human prostate cancer cell systems and human prostatic stroma via caspase-3

J Urol. 2004 Jul;172(1):338-44. doi: 10.1097/01.ju.0000132367.02834.41.


Purpose: 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.

Materials and 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Inflammatory Agents, Non-Steroidal / pharmacology*
  • Apoptosis / drug effects*
  • Aspirin / analogs & derivatives*
  • Aspirin / pharmacology*
  • Caspase 3
  • Caspases / physiology*
  • Cell Division / drug effects
  • Coloring Agents
  • Dose-Response Relationship, Drug
  • Flavanones / chemical synthesis
  • Flavanones / toxicity
  • Flow Cytometry
  • Humans
  • Ibuprofen / analogs & derivatives*
  • Ibuprofen / pharmacology*
  • In Situ Nick-End Labeling
  • Male
  • Nitrates / pharmacology*
  • Nitric Oxide Donors / pharmacology*
  • Prostatic Neoplasms / drug therapy*
  • Tetrazolium Salts
  • Thiazoles
  • Tumor Cells, Cultured


  • 2-(acetyloxy)benzoic acid 6-(nitrooxymethyl)-2-phenylmethyl ester
  • 6-(1,1-dimethylallyl)naringenin
  • Anti-Inflammatory Agents, Non-Steroidal
  • Coloring Agents
  • Flavanones
  • N-acetyl-S-(alpha-methyl-4-(2-methylpropyl)benzeneacetyl)cysteine 4-(nitrooxy)butyl ester
  • Nitrates
  • Nitric Oxide Donors
  • Tetrazolium Salts
  • Thiazoles
  • CASP3 protein, human
  • Caspase 3
  • Caspases
  • thiazolyl blue
  • Aspirin
  • Ibuprofen