Non-oxidative Loss of Glutathione in Apoptosis via GSH Extrusion

Ghibelli, Lina; Coppola, Simona; Rotilio, Giuseppe; Lafavia, E.; Maresca, Vittoria; Ciriolo, Maria Rosa

doi:10.1006/bbrc.1995.2626

Cited by 180 publications

(121 citation statements)

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“…In fact, cisplatin treatment induces a dramatic GSH depletion in frankly apoptotic cells, while such depletion is much less evident in still adhering cells at 48 h and even absent at 24 h (Table 1). A marked depletion of intracellular GSH has been described in a number of different models of apoptosis (Ghibelli et al, 1995;Macho et al, 1997), where GSH loss is consequent to an accelerated efflux via specific GSH carriers (Van den Dobbelsteen et al, 1996;Ghibelli et al, 1998;Trompier et al, 2004 To confirm calpain activation in cisplatin-treated cells and to gain information about the population of frankly apoptotic cells, we carried out the same test in intact cells using a fluorescent substrate: N-succ-Leu-Tyr-AMC (Succ) (Figure 1b). We obtained very similar results, confirming a significant increase of calpain activity at 48 h in still adhering cells, which is even more evident in detached cells.…”

Section: Calpain Activity Is Increased Early In Preapoptotic Cellsmentioning

confidence: 99%

Cross-talk between calpain and caspase-3/-7 in cisplatin-induced apoptosis of melanoma cells: a major role of calpain inhibition in cell death protection and p53 status

et al. 2006

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Section: Calpain Activity Is Increased Early In Preapoptotic Cellsmentioning

confidence: 99%

Cross-talk between calpain and caspase-3/-7 in cisplatin-induced apoptosis of melanoma cells: a major role of calpain inhibition in cell death protection and p53 status

et al. 2006

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“…In addition, alterations in its concentration have been demonstrated as a common feature of many pathological situations including AIDS, neurodegenerative diseases, and cancer (9,10). Intracellular GSH loss is an early hallmark in the progression of cell death in response to different apoptotic stimuli (11)(12)(13)(14), and has been associated with the activation of a plasma membrane transport mechanism rather than to its oxidation (11)(12)(13)(15)(16)(17). Furthermore, several studies have shown a correlation between cellular GSH depletion and the progression of apoptosis (12,13).…”

mentioning

confidence: 99%

Glutathione Depletion Is Necessary for Apoptosis in Lymphoid Cells Independent of Reactive Oxygen Species Formation

Franco

Panayiotidis²,

Cidlowski

2007

Journal of Biological Chemistry

242

193

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Changes in the intracellular redox environment of cells have been reported to be critical for the activation of apoptotic enzymes and the progression of programmed cell death. Glutathione (GSH) depletion is an early hallmark observed in apoptosis, and we have demonstrated that GSH efflux during death receptor-mediated apoptosis occurs via a GSH transporter. We now evaluate the relationship between GSH depletion, the generation of reactive oxygen species (ROS), and the progression of apoptosis. Simultaneous single cell analysis of changes in GSH content and ROS formation by multiparametric FACS revealed that loss of intracellular GSH was paralleled by the generation of different ROS including hydrogen peroxide, superoxide anion, hydroxyl radical, and lipid peroxides. However, inhibition of ROS formation by a variety of antioxidants showed that GSH loss was independent from the generation of ROS. Furthermore, GSH depletion was observed to be necessary for ROS generation. Interestingly, high extracellular thiol concentration (GSH and N-acetyl-cysteine) inhibited apoptosis, whereas, inhibition of ROS generation by other non-thiol antioxidants was ineffective in preventing cell death. Finally, GSH depletion was shown to be a necessary for the progression of apoptosis activated by both extrinsic and intrinsic signaling pathways. These results document a necessary and critical role for GSH loss in apoptosis and clearly uncouple for the first time GSH depletion from ROS formation.Apoptosis or programmed cell death is a ubiquitous, evolutionary conserved process. Under physiological conditions it is important not only in the turnover of cells in all tissues, but also during the normal development and senescence of the organism. Moreover, its deregulation has been observed to occur as either a cause or consequence of distinct pathologies (1, 2). For example, apoptosis mediated by Fas ligand (FasL) 3 receptor (Fas/CD95/Apo-1) activation is necessary for the immune system homeostasis because of its role in the rapid clearance of immunoreactive T cells maintaining the immune balance and reducing the risk of autoimmune diseases (3). Apoptosis is a highly organized process characterized by the progressive activation of precise pathways leading to specific biochemical and morphological alterations. Initial stages of apoptosis are characterized by reactive oxygen species (ROS) formation, changes in intracellular ionic homeostasis, cell shrinkage, loss of membrane lipid asymmetry, and chromatin condensation. Later stages associated with the execution phase of apoptosis are characterized by activation of execution caspases and endonucleases, apoptotic body formation, and cell fragmentation (1, 4, 5). Apoptosis has been widely reported to be modulated by changes in the redox status of the cell; however, the precise mechanisms involved are still unclear (6, 7). Reduced glutathione (GSH) is the most abundant low molecular weight thiol in animal cells and is involved in many cellular processes including antioxidant defense, drug det...

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“…On the other hand, ROS also have an important function in pro-apoptotic pathways (Curtin et al, 2002;Matsuzawa and Ichijo, 2005). As intracellular redox status is critical in determining the survival and death of cells (Hampton and Orrenius, 1998), many cancer therapeutics target to induce cellular apoptosis by disrupting the redox balance and depleting the intracellular thiol buffer system through extrusion or redistribution of glutathione (GSH) (Ghibelli et al, 1995).…”

mentioning

confidence: 99%

Role of reactive oxygen species in brucein D-mediated p38-mitogen-activated protein kinase and nuclear factor-κB signalling pathways in human pancreatic adenocarcinoma cells

2010

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BACKGROUND: In human pancreatic adenocarcinoma, nuclear factor-kappa-B (NF-kB) transcription factor is constitutively activated that contributes to the resistance of the tumour cells to induced apoptosis. In our earlier studies, we have shown that brucein D (BD) mediated apoptosis through activation of the p38-mitogen-activated protein kinase (MAPK) signalling pathway in pancreatic cancer cells. This study investigated the function of reactive oxygen species (ROS) in BD-mediated p38-MAPK and NF-kB signalling pathways in PANC-1 cells. METHODS: Glutathione and dihydroethidium assays were used to measure the antioxidant and superoxide levels, respectively. The protein expression of p22 phox , p67 phox and p38-MAPK were examined by western blot. The NF-kB activity was evaluated by electrophoretic mobility shift assay. RESULTS: Treatment with BD depleted the intracellular glutathione levels in PANC-1 cells. Brucein D triggered the activation of NADPH oxidase isoforms, p22 phox and p67 phox while enhancing the generation of superoxide. Increases in both intracellular ROS and NADPH oxidase activity were inhibited by an antioxidant, N-acetylcysteine (NAC). Brucein D-mediated activation of p38-MAPK was also inhibited by NAC. However, inhibition of NF-kB activity in BD-treated cells was independent of ROS. In vivo studies showed that BD treatment effectively reduced the rate of xenograft human pancreatic tumour in nude mice with no significant toxicity. CONCLUSION: These data suggest that BD is an apoptogenic agent for pancreatic cancer cells through activation of the redox-sensitive p38-MAPK pathway and inhibition of NF-kB anti-apoptotic activity in pancreatic cancer cells.

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Non-oxidative Loss of Glutathione in Apoptosis via GSH Extrusion

Cited by 180 publications

References 0 publications

Cross-talk between calpain and caspase-3/-7 in cisplatin-induced apoptosis of melanoma cells: a major role of calpain inhibition in cell death protection and p53 status

Cross-talk between calpain and caspase-3/-7 in cisplatin-induced apoptosis of melanoma cells: a major role of calpain inhibition in cell death protection and p53 status

Glutathione Depletion Is Necessary for Apoptosis in Lymphoid Cells Independent of Reactive Oxygen Species Formation

Role of reactive oxygen species in brucein D-mediated p38-mitogen-activated protein kinase and nuclear factor-κB signalling pathways in human pancreatic adenocarcinoma cells

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