Nitric oxide donating nonsteroidal anti-inflammatory drugs induce apoptosis in human prostate cancer cell systems and human prostatic stroma via caspase-3.
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.
Royle
Justine Sarah
JS
Department of Urology, Western General Hospital, Prostate Research Group, University of Edinburgh, Western General Hospital, Edinburgh, United Kingdom. justine.royle@virgin.net
Ross
James A
JA
Ansell
Ian
I
Bollina
Prasad
P
Tulloch
David N
DN
Habib
Fouad K
FK
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
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