scholarly journals Zinc Chelation Mediates the Lysosomal Disruption without Intracellular ROS Generation

2016 ◽  
Vol 2016 ◽  
pp. 1-13 ◽  
Author(s):  
Andreza Cândido Matias ◽  
Tânia Maria Manieri ◽  
Giselle Cerchiaro
Keyword(s):  
Cell Death ◽  
Molecular Mechanism ◽  
Caspase 3 ◽  
Neuroblastoma Cells ◽  
P38 Map Kinase ◽  
Ros Generation ◽  
Intracellular Zinc ◽  
Intracellular Ros ◽  
Zinc Depletion ◽  
Zinc Chelation

We report the molecular mechanism for zinc depletion caused by TPEN (N,N,N′,N′-Tetrakis(2-pyridylmethyl)ethylenediamine) in neuroblastoma cells. The activation of p38 MAP kinase and subsequently caspase 3 is not due to or followed by redox imbalance or ROS generation, though these are commonly observed in literature. We found that TPEN is not responsible for ROS generation and the mechanism involves essentially lysosomal disruption caused by intracellular zinc depletion. We also observed a modest activation of Bax and no changes in the Bcl-2 proteins. As a result, we suggest that TPEN causes intracellular zinc depletion which can influence the breakdown of lysosomes and cell death without ROS generation.

scholarly journals Generation of Cellular Reactive Oxygen Species by Activation of the EP2 Receptor Contributes to Prostaglandin E2-Induced Cytotoxicity in Motor Neuron-Like NSC-34 Cells

2020 ◽  
Vol 2020 ◽  
pp. 1-14 ◽  
Author(s):  
Yasuhiro Kosuge ◽  
Hiroshi Nango ◽  
Hiroki Kasai ◽  
Takuya Yanagi ◽  
Takayuki Mawatari ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Cell Death ◽  
Prostaglandin E2 ◽  
Motor Neuron ◽  
Motor Neurons ◽  
Caspase 3 ◽  
Reactive Oxygen ◽  
Ros Generation ◽  
Oxygen Species ◽  
Intracellular Ros

Amyotrophic lateral sclerosis (ALS) is a devastating motor neuron disease characterized by progressive degeneration of motor neurons in the central nervous system. Prostaglandin E2 (PGE2) plays a pivotal role in the degeneration of motor neurons in human and transgenic models of ALS. We have shown previously that PGE2 directly induces neuronal death through activation of the E-prostanoid (EP) 2 receptor in differentiated NSC-34 cells, a motor neuron-like cell line. In the present study, to clarify the mechanisms underlying PGE2-induced neurotoxicity, we focused on generation of intracellular reactive oxygen species (ROS) and examined the effects of N-acetylcysteine (NAC), a cell-permeable antioxidant, on PGE2-induced cell death in differentiated NSC-34 cells. Dichlorofluorescein (DCF) fluorescence analysis of PGE2-treated cells showed that intracellular ROS levels increased markedly with time, and that this effect was antagonized by a selective EP2 antagonist (PF-04418948) but not a selective EP3 antagonist (L-798,106). Although an EP2-selective agonist, butaprost, mimicked the effect of PGE2, an EP1/EP3 agonist, sulprostone, transiently but significantly decreased the level of intracellular ROS in these cells. MTT reduction assay and lactate dehydrogenase release assay revealed that PGE2- and butaprost-induced cell death were each suppressed by pretreatment with NAC in a concentration-dependent manner. Western blot analysis revealed that the active form of caspase-3 was markedly increased in the PGE2- and butaprost-treated cells. These increases in caspase-3 protein expression were suppressed by pretreatment with NAC. Moreover, dibutyryl-cAMP treatment of differentiated NSC-34 cells caused intracellular ROS generation and cell death. Our data reveal the existence of a PGE2-EP2 signaling-dependent intracellular ROS generation pathway, with subsequent activation of the caspase-3 cascade, in differentiated NSC-34 cells, suggesting that PGE2 is likely a key molecule linking inflammation to oxidative stress in motor neuron-like NSC-34 cells.

scholarly journals 1-Methyl-4-phenylpyridinium (MPP+)-induced apoptosis and mitochondrial oxidant generation: role of transferrin-receptor-dependent iron and hydrogen peroxide

2003 ◽  
Vol 371 (1) ◽  
pp. 151-164 ◽  
Author(s):  
Shasi V. KALIVENDI ◽  
Srigiridhar KOTAMRAJU ◽  
Sonya CUNNINGHAM ◽  
Tiesong SHANG ◽  
Cecilia J. HILLARD ◽  
...  
Keyword(s):  
Cell Death ◽  
Transferrin Receptor ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Neuroblastoma Cells ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Ros Generation ◽  
Cellular Models ◽  
Pre Treatment ◽  
Induced Apoptosis

1-Methyl-4-phenylpyridinium (MPP+) is a neurotoxin used in cellular models of Parkinson's Disease. Although intracellular iron plays a crucial role in MPP+-induced apoptosis, the molecular signalling mechanisms linking iron, reactive oxygen species (ROS) and apoptosis are still unknown. We investigated these aspects using cerebellar granule neurons (CGNs) and human SH-SY5Y neuroblastoma cells. MPP+ enhanced caspase 3 activity after 24h with significant increases as early as 12h after treatment of cells. Pre-treatment of CGNs and neuroblastoma cells with the metalloporphyrin antioxidant enzyme mimic, Fe(III)tetrakis(4-benzoic acid)porphyrin (FeTBAP), completely prevented the MPP+-induced caspase 3 activity as did overexpression of glutathione peroxidase (GPx1) and pre-treatment with a lipophilic, cell-permeable iron chelator [N,N′-bis-(2-hydroxybenzyl)ethylenediamine-N,N′-diacetic acid, HBED]. MPP+ treatment increased the number of TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labelling)-positive cells which was completely blocked by pre-treatment with FeTBAP. MPP+ treatment significantly decreased the aconitase and mitochondrial complex I activities; pre-treatment with FeTBAP, HBED and GPx1 overexpression reversed this effect. MPP+ treatment increased the intracellular oxidative stress by 2—3-fold, as determined by oxidation of dichlorodihydrofluorescein and dihydroethidium (hydroethidine). These effects were reversed by pre-treatment of cells with FeTBAP and HBED and by GPx1 overexpression. MPP+-treatment enhanced the cell-surface transferrin receptor (TfR) expression, suggesting a role for TfR-induced iron uptake in MPP+ toxicity. Treatment of cells with anti-TfR antibody (IgA class) inhibited MPP+-induced caspase activation. Inhibition of nitric oxide synthase activity did not affect caspase 3 activity, apoptotic cell death or ROS generation by MPP+. Overall, these results suggest that MPP+-induced cell death in CGNs and neuroblastoma cells proceeds via apoptosis and involves mitochondrial release of ROS and TfR-dependent iron.

p38 MAP Kinase Mediates the Cell Death Induced by PrP106–126 in the SH-SY5Y Neuroblastoma Cells

2002 ◽  
Vol 9 (1) ◽  
pp. 69-81 ◽  
Author(s):  
Stefano Thellung ◽  
Valentina Villa ◽  
Alessandro Corsaro ◽  
Sara Arena ◽  
Enrico Millo ◽  
...  
Keyword(s):  
Cell Death ◽  
Map Kinase ◽  
Neuroblastoma Cells ◽  
P38 Map Kinase

Honokiol Induces Autophagic Apoptosis in Neuroblastoma Cells through a P53-Dependent Pathway

2019 ◽  
Vol 47 (04) ◽  
pp. 895-912 ◽  
Author(s):  
Ming-Chung Lin ◽  
Yuan-Wen Lee ◽  
Yuan-Yun Tseng ◽  
Yung-Wei Lin ◽  
Jui-Tai Chen ◽  
...  
Keyword(s):  
Dna Damage ◽  
Cell Death ◽  
Cell Apoptosis ◽  
Caspase 3 ◽  
Therapeutic Option ◽  
Neuroblastoma Cells ◽  
Cytochrome C Release ◽  
Dependent Pathway ◽  
Dependent Mechanism ◽  
Stimulation Of

In children, neuroblastomas are the most common and deadly solid tumor. Our previous studies showed that honokiol can cross the blood–brain barrier and kill neuroblastoma cells. In this study, we further evaluated if exposure to honokiol for short periods could induce autophagy and subsequent apoptosis of neuroblastoma cells and possible mechanisms. Exposure of neuroblastoma neuro-2a cells to honokiol for 24[Formula: see text]h induced morphological shrinkage and cell death. As to the mechanisms, honokiol consecutively induced cytochrome c release from mitochondria, caspase-3 activation, DNA fragmentation and cell apoptosis. Separately, honokiol time-dependently augmented the proportion of autophagic cells and the ratio of light chain 3 (LC3)-II/LC3-I. Pretreatment of neuro-2a cells with 3-methyladenine, an inhibitor of autophagy, attenuated honokiol-induced cell autophagy, caspase-3 activation, DNA damage and cell apoptosis. In contrast, stimulation of autophagy by rapamycin, an inducer of autophagy, significantly enhanced honokiol-induced cell apoptosis. Furthermore, honokiol-induced autophagic apoptosis was confirmed in neuroblastoma NB41A3 cells. Knocking down translation of p53 using RNA interference attenuated honokiol-induced autophagy and apoptosis in neuro-2a and NB41A3 cells. Taken together, this study showed that at early periods, honokiol can induce autophagic apoptosis of neuroblastoma cells through activating a p53-dependent mechanism. Consequently, honokiol has the potential to be a therapeutic option for neuroblastomas.

Clinical and Molecular Significance of Tumor Necrosis In Newly Diagnosed Patients with Hodgkin's Lymphoma and Diffuse Large B Cell Lymphoma

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1995-1995
Author(s):  
Ihab Abd-Elrahman ◽  
Vladimir Rapoport ◽  
Tzahi Neuman ◽  
Liat Appelbaum ◽  
Tamar Shiloach ◽  
...  
Keyword(s):  
Cell Death ◽  
Molecular Mechanism ◽  
Tumor Necrosis ◽  
Caspase 3 ◽  
B Cell Lymphoma ◽  
Newly Diagnosed ◽  
Ki 67 ◽  
Bulky Disease ◽  
Radiographic Appearance ◽  
Tumor Mass

Abstract Abstract 1995 Lymphoma of various types in newly diagnosed patients may exhibit necrotic areas in the tumor mass on imaging. To date, little is known about the clinical significance of this finding and even less is known about the mechanism of cell death in the necrotic tissue - apoptosis, molecular necrosis or autophagy. The objective of this study was to investigate the prognostic significance of tumor necrosis in newly diagnosed Hodgkin's lymphoma (HL) and diffuse large B cell lymphoma (DLBCL) patients and to define the molecular mechanism of cell death responsible for the necrosis. Two hundred and four CT scan or CT-PET studies from newly diagnosed patients with DLBCL (131) and HL (73) were analyzed for the presence of tumor necrosis. Radiographic appearance of necrosis was present in 47% of all patients, 34% HL and 53% DLBCL. A statistically significant correlation was found between necrosis and bulky disease (tumor mass size≥10 cm) (p=0.0002×10-6) and also between necrosis and elevated LDH (p=0.00002) (see Table 1). Radiographic appearance of necrosis Bulky disease tumor mass size ≥10 cm Elevated LDH Yes 41.1% 60.2% No 5.5% 29.9% No statistically significant correlation was found between necrosis and age >60 or more advanced stage of disease. There was also no statistically significant difference in interim response to treatment, disease-free survival and overall survival between patients with necrosis and without it. Our results show that in contrast to solid tumors, tissue necrosis in lymphoma is not a predictor of worse prognosis: though more patients with radiographic appearance of necrosis had also the known bad prognostic signs- elevated LDH and/or bulky disease, this group did not have worse outcome. We suggest that necrosis can lead to a better response to treatment in lymphoma. Among 56 HL and DLBCL patients that underwent an open surgical biopsy of the tumor mass nineteen presented necrotic morphology in the biopsied tissues: 8 HL and 11 DLBCL. Biopsies without evidence of necrotic morphology (10 patients from the above 56) and reactive lymph nodes (4) were included as controls. All sections were stained for ki-67 (proliferation marker), activated caspase-3 (apoptosis marker) and HMGB-1 (necrotic cell death marker). DLBCL cells showed high proliferation rate, with ki-67 index ranging from 40 to 90% and in HL cells the ki-67 index ranged from 5 to 50%. All the samples positive for HMGB-1 were from patients with HL and 10/11 of activated caspase-3 positive samples were from DLBCL. These results show a differential molecular mechanism of cell death in the tumor mass of newly diagnosed HL and DLBCL patients. The constitutive activation of the survival factor Nuclear Factor-kB (NF-κB) was found in Hodgkin and non-Hodgkin lymphoma. We stained the same biopsies for p65/RelA and examined the nuclear localization of NF-κB as an indication for activation of the NF-κB pathway. Four DLBCL and 4 HL samples were positive for NF-κB activity. Nevertheless, we found that activation of caspase-3 and/or HMGB-1 induced cell death in these tissues. In summary, tumor necrosis in HL and DLBCL is correlated with bulky disease and high LDH, yet is not correlated with prognosis. The molecular mechanism of cell death differs between the two types of lymphoma. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Bufalin Induces Reactive Oxygen Species Dependent Bax Translocation and Apoptosis in ASTC-a-1 Cells

2011 ◽  
Vol 2011 ◽  
pp. 1-12 ◽  
Author(s):  
Lei Sun ◽  
Tongsheng Chen ◽  
Xiaoping Wang ◽  
Yun Chen ◽  
Xunbin Wei
Keyword(s):  
Reactive Oxygen Species ◽  
Cell Death ◽  
Molecular Mechanisms ◽  
Caspase 3 ◽  
Temporal Dynamics ◽  
Reactive Oxygen ◽  
Ros Generation ◽  
Ros Production ◽  
Oxygen Species ◽  
Induced Apoptosis

Bufalin has been shown to induce cancer cell death through apoptotic pathways. However, the molecular mechanisms are not well understood. In this study, we used the confocal fluorescence microscopy (CFM) to monitor the spatio-temporal dynamics of reactive oxygen species (ROS) production, Bax translocation and caspase-3 activation during bufalin-induced apoptosis in living human lung adenocarcinoma (ASTC-a-1) cells. Bufalin induced ROS production and apoptotic cell death, demonstrated by Hoechst 33258 staining as well as flow cytometry analysis. Bax redistributed from cytosol to mitochondria from 12 to 48 h after bufalin treatment in living cells expressed with green fluorescent protein Bax. Treatment with the antioxidantN-acetyl-cysteine (NAC), a ROS scavenger, inhibited ROS generation and Bax translocation and led to a significant protection against bufalin-induced apoptosis. Our results also revealed that bufalin induced a prominent increase of caspase-3 activation blocked potently by NAC. Taken together, bufalin induced ROS-mediated Bax translocation, mitochondrial permeability transition and caspase-3 activation, implying that bufalin induced apoptosis via ROS-dependent mitochondrial death pathway in ASTC-a-1 cells.

Citicoline increases glutathione redox ratio and reduces caspase-3 activation and cell death in staurosporine-treated SH-SY5Y human neuroblastoma cells

2002 ◽  
Vol 957 (1) ◽  
pp. 84-90 ◽  
Author(s):  
Marta Barrachina ◽  
Julio Secades ◽  
Rafael Lozano ◽  
Cristina Gómez-Santos ◽  
Santiago Ambrosio ◽  
...  
Keyword(s):  
Cell Death ◽  
Caspase 3 ◽  
Neuroblastoma Cells ◽  
Human Neuroblastoma Cells ◽  
Human Neuroblastoma ◽  
Redox Ratio ◽  
Glutathione Redox ◽  
Glutathione Redox Ratio

scholarly journals Neuroblastoma Cell Death Induced by eEF1A2 Knockdown Is Possibly Mediated by the Inhibition of Akt and mTOR Phosphorylation

2021 ◽  
Author(s):  
Kawinthra Khwanraj ◽  
Permphan Dharmasaroja
Keyword(s):  
Cell Death ◽  
Caspase 3 ◽  
Neuroblastoma Cell ◽  
Neuroblastoma Cells ◽  
Elongation Factor ◽  
Mtor Pathway ◽  
Eukaryotic Translation ◽  
Mtor Phosphorylation ◽  
Almost All

Background The protein kinase B/mammalian target of the rapamycin (Akt/mTOR) pathway is one of the most potent prosurvival signaling cascades that is constitutively active in neuroblastoma. The eukaryotic translation elongation factor-1, alpha-2 (eEF1A2) protein has been found to activate the Akt/mTOR pathway. However, there is a lack of data on the role of eEF1A2 in neuroblastoma. The present study investigated the effect of eEF1A2 silencing on the viability of neuroblastoma cells and its possible signaling. Materials and Methods: Human SH-SY5Y neuroblastoma cells were transfected with small interfering RNA (siRNA) against eEF1A2. After 48 h of transfection, cell viability was assessed using an MTT assay. The mRNA expression of p53, Bax, Bcl-2, caspase-3 and members of the phosphoinositide 3-kinases (PI3K)/Akt/mTOR pathway was determined using quantitative real-time RT-PCR (qRT-PCR). The protein expression of Akt and mTOR was measured using Western blot analysis. Results: eEF1A2 knockdown significantly decreased the viability of neuroblastoma cells. No significant changes were observed on the expression of p53, Bax/Bcl-2 ratio, and caspase-3 mRNAs; however, the upregulated trends were noted for the p53 and Bax/Bcl-2 ratio. eEF1A2 knockdown significantly inhibited the phosphorylation of both Akt and mTOR. Almost all of the class I (PIK3CA, PIK3CB, and PIK3CD) and all of the class II PI3K genes were slightly increased in tumor cells with eEF1A2 knockdown. In addition, a slightly decreased expression of the Akt2, mTORC1, and mTORC2 was observed. Conclusion: eEF1A2 knockdown induced neuroblastoma cell death, in part through the inhibition of Akt and mTOR, suggesting a potential role of eEF1A2 as a molecular target for neuroblastoma therapy.

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