Nitric oxide-releasing prodrug triggers cancer cell death through deregulation of cellular redox balance.
Redox Biol
; 1: 115-24, 2013.
Article
em En
| MEDLINE
| ID: mdl-24024144
ABSTRACT
JS-K is a nitric oxide (NO)-releasing prodrug of the O (2)-arylated diazeniumdiolate family that has demonstrated pronounced cytotoxicity and antitumor properties in a variety of cancer models both in vitro and in vivo. The current study of the metabolic actions of JS-K was undertaken to investigate mechanisms of its cytotoxicity. Consistent with model chemical reactions, the activating step in the metabolism of JS-K in the cell is the dearylation of the diazeniumdiolate by glutathione (GSH) via a nucleophilic aromatic substitution reaction. The resulting product (CEP/NO anion) spontaneously hydrolyzes, releasing two equivalents of NO. The GSH/GSSG redox couple is considered to be the major redox buffer of the cell, helping maintain a reducing environment under basal conditions. We have quantified the effects of JS-K on cellular GSH content, and show that JS-K markedly depletes GSH, due to JS-K's rapid uptake and cascading release of NO and reactive nitrogen species. The depletion of GSH results in alterations in the redox potential of the cellular environment, initiating MAPK stress signaling pathways, and inducing apoptosis. Microarray analysis confirmed signaling gene changes at the transcriptional level and revealed alteration in the expression of several genes crucial for maintenance of cellular redox homeostasis, as well as cell proliferation and survival, including MYC. Pre-treating cells with the known GSH precursor and nucleophilic reducing agent N-acetylcysteine prevented the signaling events that lead to apoptosis. These data indicate that multiplicative depletion of the reduced glutathione pool and deregulation of intracellular redox balance are important initial steps in the mechanism of JS-K's cytotoxic action.
Palavras-chave
ATF, activating transcription factor; Arylated diazeniumdiolate; DAF-FM, 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate; DCF-DA, 5-(and 6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate; DMSO, dimethyl sulfoxide; FBS, fetal bovine serum; GSH, glutathione; GSSG, glutathione disulfide (oxidized GSH); Glutathione; HBSS, Hank's balanced salt solution; IPA, Ingenuity Pathway Analysis; JS-K, O2-(2,4-dinitrophenyl) 1-[(4-ethoxycarbonyl)piperazin-1-yl]diazen-1-ium-1,2-diolate; LC/MS, liquid chromatography/mass spectrometry; Leukemia; MAPK, mitogen-activated protein kinase; NAC, N-acetylcysteine; NO, nitric oxide; NSCLC, non-small cell lung cancer; Nitric oxide; PARP, poly (ADP-ribose) polymerase; RNS, reactive nitrogen species; ROS, reactive oxygen species; SAPK/JNK, stress activated protein kinase/c-jun N-terminal kinase.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Piperazinas
/
Compostos Azo
/
Pró-Fármacos
/
Leucemia
/
Doadores de Óxido Nítrico
Tipo de estudo:
Prognostic_studies
Limite:
Humans
Idioma:
En
Revista:
Redox Biol
Ano de publicação:
2013
Tipo de documento:
Article
País de afiliação:
Estados Unidos