title: NO-releasing NSAIDs suppress NF-κB signaling in vitro and in vivo through S-nitrosylation.
abstract: NO-NSAIDs are promising anticancer drugs, comprising an NSAID, an NO-releasing moiety, and a spacer linking them. Although the effect of NO-NSAIDs on a wide variety of signaling and other cellular mechanisms has been deciphered, a key question remains unanswered, that being the role of NO to the overall biological effect of these agents. It has been shown that NO can directly modify sulfhydryl residues of proteins through S-nitrosylation and induce apoptosis. We studied 3 NO-NSAIDs having a different NSAID, spacer, and NO-releasing moiety. In vitro: aspirin, NO-ASA, naproxen, and NO-naproxen inhibited HT-29 human colon cancer cell growth, the IC(50)s being >5000, 192±6, 2800±210 and 95±5μM at 24h, respectively. NO-Aspirin and NO-naproxen reduced NF-κB protein levels, and activated caspase-3 enzyme in a dose- and time-dependent manner. Based on the biotin switch assay, NO-ASA and NO-naproxen S-nitrosylated NF-κB p65 in a time-dependent manner. Pretreatment of the cells with carboxy-PTIO, abrogated the S-nitrosylation of NF-κB p65. In vivo: rats treated with NO-ASA, NONO-ASA, and NO-naproxen showed S-nitrosylation of NF-κB p65 in the stomach tissue, increases in plasma TNF-α, and reductions in mucosal PGE(2) levels. These data provide a mechanistic role for NO and a rational for the chemopreventive effects of NO-NSAIDs.
authors.affiliation: Department of Physiology and Pharmacology, City University of New York Medical School, NY 10031, USA.
authors.forename: Mitali
authors.initials: M
authors.lastname: Chattopadhyay
authors.forename: Satindra
authors.initials: S
authors.lastname: Goswami
authors.forename: Deborah B
authors.initials: DB
authors.lastname: Rodes
authors.forename: Ravinder
authors.initials: R
authors.lastname: Kodela
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Velazquez
authors.forename: Daniel
authors.initials: D
authors.lastname: Boring
authors.forename: James A
authors.initials: JA
authors.lastname: Crowell
authors.forename: Khosrow
authors.initials: K
authors.lastname: Kashfi
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism / Q000494:pharmacology*; D001241:Aspirin / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D015153:Blotting, Western; D053148:Caspase 3 / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D003110:Colonic Neoplasms / Q000378:metabolism / Q000473:pathology; D015232:Dinoprostone / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005753:Gastric Mucosa / Q000187:drug effects / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D008297:Male; D016328:NF-kappa B / Q000378:metabolism*; D009288:Naproxen / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D009566:Nitrates / Q000378:metabolism / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism; D020030:Nitric Oxide Donors / Q000378:metabolism / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D026403:S-Nitrosothiols / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D013270:Stomach / Q000187:drug effects; D051996:Transcription Factor RelA / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000097:blood
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; C488176:N-acetyl-S-(alpha-methyl-4-(2-methylpropyl)benzeneacetyl)cysteine 4-(nitrooxy)butyl ester; D016328:NF-kappa B; D009566:Nitrates; D020030:Nitric Oxide Donors; D026403:S-Nitrosothiols; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; C111751:naproxen-n-butyl nitrate; C098312:nitroxy-butyl-acetylsalicylic acid; D009569:Nitric Oxide; D009288:Naproxen; D053148:Caspase 3; D015232:Dinoprostone; D001241:Aspirin