Pro-apoptotic effect of a nitric oxide-donating NSAID, NCX 4040, on bladder carcinoma cells.
Nitric oxide-releasing non steroidal anti-inflammatory drugs (NO-NSAIDs) are a promising class of compounds that cause cell cycle perturbations and induce apoptosis in cell lines from different tumors. We investigated the activity of a recently developed NO-NSAID (NCX 4040) in bladder cancer cell lines (HT1376 and MCR). Cells were treated with different drug concentrations for different exposure times. Cytostatic and cytocidal activity was tested by SRB assay and apoptosis was evaluated by TUNEL analysis, ANNEXIN V assay and fluorescence microscopy. To further investigate the cell death-inducing mechanisms of NCX 4040, we analyzed gp-170, caspase expression and mitochondrial membrane potential (Delta Psi) depolarization. NCX 4040 showed a striking cytocidal activity in both cell lines, reaching LC(50) at a 10-microM and 50-microM concentrations in HT1376 and in MCR cells, respectively, after an exposure of only 6 h followed by an 18-h washout. Apoptosis was triggered in up to 90% of cells and was associated with active caspase-3 expression and Delta Psi depolarization in both cell lines after a 6-h exposure. In conclusion, NCX 4040, which probably causes apoptosis via a mitochondrial-dependent mechanism, could prove to be a useful agent for improving bladder cancer treatment.
Fabbri
F
F
Istituto Oncologico Romagnolo, Morgagni-Pierantoni Hospital, Via Forlanini 34, 47100 Forlì, Italy.
Brigliadori
G
G
Ulivi
P
P
Tesei
A
A
Vannini
I
I
Rosetti
M
M
Bravaccini
S
S
Amadori
D
D
Bolla
M
M
Zoli
W
W
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology* / Q000627:therapeutic use; D000970:Antineoplastic Agents / Q000494:pharmacology / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D001241:Aspirin / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D006801:Humans; D020287:In Situ Nick-End Labeling; D008564:Membrane Potentials / Q000187:drug effects; D009569:Nitric Oxide / Q000378:metabolism*; D009574:Nitro Compounds / Q000494:pharmacology* / Q000627:therapeutic use; D012459:Salicylates / Q000494:pharmacology / Q000627:therapeutic use; D001749:Urinary Bladder Neoplasms / Q000188:drug therapy*
D000894:Anti-Inflammatory Agents, Non-Steroidal; D000970:Antineoplastic Agents; C498045:NCX 4040; C507361:NCX 4042; D009574:Nitro Compounds; D012459:Salicylates; D009569:Nitric Oxide; D001241:Aspirin