scholarly journals ATF3 contributes to brucine-triggered glioma cell ferroptosis via promotion of hydrogen peroxide and iron

2021 ◽  
Author(s):  
Shan Lu ◽  
Xuan-zhong Wang ◽  
Chuan He ◽  
Lei Wang ◽  
Shi-peng Liang ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Cell Death ◽  
Er Stress ◽  
Glioma Cell ◽  
Nuclear Translocation ◽  
Indole Alkaloid ◽  
Ammonium Citrate ◽  
Ferric Ammonium Citrate ◽  
Activating Transcription Factor 3 ◽  
Intracellular Iron

AbstractFerroptotic cell death is characterized by iron-dependent lipid peroxidation that is initiated by ferrous iron and H2O2 via Fenton reaction, in which the role of activating transcription factor 3 (ATF3) remains elusive. Brucine is a weak alkaline indole alkaloid extracted from the seeds of Strychnos nux-vomica, which has shown potent antitumor activity against various tumors, including glioma. In this study, we showed that brucine inhibited glioma cell growth in vitro and in vivo, which was paralleled by nuclear translocation of ATF3, lipid peroxidation, and increases of iron and H2O2. Furthermore, brucine-induced lipid peroxidation was inhibited or exacerbated when intracellular iron was chelated by deferoxamine (500 μM) or improved by ferric ammonium citrate (500 μM). Suppression of lipid peroxidation with lipophilic antioxidants ferrostatin-1 (50 μM) or liproxstatin-1 (30 μM) rescued brucine-induced glioma cell death. Moreover, knockdown of ATF3 prevented brucine-induced accumulation of iron and H2O2 and glioma cell death. We revealed that brucine induced ATF3 upregulation and translocation into nuclei via activation of ER stress. ATF3 promoted brucine-induced H2O2 accumulation via upregulating NOX4 and SOD1 to generate H2O2 on one hand, and downregulating catalase and xCT to prevent H2O2 degradation on the other hand. H2O2 then contributed to brucine-triggered iron increase and transferrin receptor upregulation, as well as lipid peroxidation. This was further verified by treating glioma cells with exogenous H2O2 alone. Moreover, H2O2 reversely exacerbated brucine-induced ER stress. Taken together, ATF3 contributes to brucine-induced glioma cell ferroptosis via increasing H2O2 and iron.

Regulation of xanthine oxidoreductase by intracellular iron

2002 ◽  
Vol 283 (6) ◽  
pp. C1722-C1728 ◽  
Author(s):  
Eeva Martelin ◽  
Risto Lapatto ◽  
Kari O. Raivio
Keyword(s):  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Iron Chelation ◽  
Mouse Fibroblast ◽  
Ammonium Citrate ◽  
Ferric Ammonium Citrate ◽  
Mrna Levels ◽  
Xanthine Oxidoreductase ◽  
Intracellular Iron ◽  
Human Embryonic Kidney Cells

Xanthine oxidoreductase (XOR) may produce reactive oxygen species and play a role in ischemia-reperfusion injury. Because tissue iron levels increase after ischemia, and because XOR contains functionally critical iron-sulfur clusters, we studied the effects of intracellular iron on XOR expression. Ferric ammonium citrate and FeSO4elevated intracellular iron levels and increased XOR activity up to twofold in mouse fibroblast and human bronchial epithelial cells. Iron increased XOR protein and mRNA levels, whereas protein and RNA synthesis inhibitors abolished the induction of XOR activity. A human XOR promoter construct (nucleotides +42 to −1937) was not induced by iron in human embryonic kidney cells. Hydroxyl radical scavengers did not block induction of XOR activity by iron. Iron chelation by deferoxamine (DFO) decreased XOR activity but did not lower endogenous XOR protein or mRNA levels. Furthermore, DFO reduced the activity of overexpressed human XOR but not the amount of immunoreactive protein. Our data show that XOR activity is transcriptionally induced by iron but posttranslationally inactivated by iron chelation.

scholarly journals Iron Causes Lipid Oxidation and Inhibits Proteasome Function in Multiple Myeloma Cells: A Proof of Concept for Novel Combination Therapies

Cancers ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 970 ◽  
Author(s):  
Jessica Bordini ◽  
Federica Morisi ◽  
Fulvia Cerruti ◽  
Paolo Cascio ◽  
Clara Camaschella ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Cell Death ◽  
Cell Lines ◽  
Iron Toxicity ◽  
Ammonium Citrate ◽  
Ferric Ammonium Citrate ◽  
Ferric Carboxymaltose ◽  
High Dose ◽  
Iron Loading

Adaptation to import iron for proliferation makes cancer cells potentially sensitive to iron toxicity. Iron loading impairs multiple myeloma (MM) cell proliferation and increases the efficacy of the proteasome inhibitor bortezomib. Here, we defined the mechanisms of iron toxicity in MM.1S, U266, H929, and OPM-2 MM cell lines, and validated this strategy in preclinical studies using Vk*MYC mice as MM model. High-dose ferric ammonium citrate triggered cell death in all cell lines tested, increasing malondialdehyde levels, the by-product of lipid peroxidation and index of ferroptosis. In addition, iron exposure caused dose-dependent accumulation of polyubiquitinated proteins in highly iron-sensitive MM.1S and H929 cells, suggesting that proteasome workload contributes to iron sensitivity. Accordingly, high iron concentrations inhibited the proteasomal chymotrypsin-like activity of 26S particles and of MM cellular extracts in vitro. In all MM cells, bortezomib-iron combination induced persistent lipid damage, exacerbated bortezomib-induced polyubiquitinated proteins accumulation, and triggered cell death more efficiently than individual treatments. In Vk*MYC mice, addition of iron dextran or ferric carboxymaltose to the bortezomib-melphalan-prednisone (VMP) regimen increased the therapeutic response and prolonged remission without causing evident toxicity. We conclude that iron loading interferes both with redox and protein homeostasis, a property that can be exploited to design novel combination strategies including iron supplementation, to increase the efficacy of current MM therapies.

scholarly journals NCOA4-Mediated Ferritinophagy Is Essential for HT22 Neuronal Cell Survival During Iron Deficiency (OR15-07-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Emily Bengson ◽  
Moon-Suhn Ryu
Keyword(s):  
Cell Death ◽  
Iron Deficiency ◽  
Iron Overload ◽  
Cell Survival ◽  
Neuronal Cell ◽  
Ammonium Citrate ◽  
Ferric Ammonium Citrate ◽  
Mrna Levels ◽  
Ht22 Cells ◽  
Cellular Iron

Abstract Objectives Iron is essential for proper cell function and development. However, mishandled iron may lead to ferroptotic cell death. NCOA4 manages the cellular labile iron pool by controlling the release of ferritin iron via ferritinophagy. The present studies examined the capacity of hippocampal cells in handling iron fluctuation, with particular focus on NCOA4 and ferritin. Methods HT22 mouse hippocampal cells were treated with ferric ammonium citrate (FAC) and deferoxamine (Dfo) to produce cellular iron overload and deprivation, respectively. Ferroptosis was determined by measures of ferrostatin-1 effects and Ptgs2 mRNA. For ferritinophagy studies, Ncoa4 was silenced by siRNA transfections. Functional impacts of impaired ferritinophagy were assessed via CCK-8 cell viability assays and western and qPCR analyses of iron-related genes. Results HT22 cells were highly susceptible to cellular iron overload. FAC-treated cells featured acute morphological changes, decreased viability, and elevated Ptgs2 mRNA abundance. Iron effects were prevented by ferrostatin-1, indicating ferroptosis by cellular iron overload. Dfo alone had minimal impact on cell morphology and viability. NCOA4 protein, but not mRNA, levels were acutely upregulated by Dfo treatments. Ferritin turnover by iron deficiency was impaired in NCOA4-depleted cells, presumably due to impaired ferritinophagy. Moreover, HT22 cells became sensitive to iron deficiency by loss of NCOA4. Conclusions Our studies demonstrate iron can induce ferroptosis in neuronal cells. We also identify NCOA4-mediated ferritinophagy as an integral process for neuronal cell survival during iron deficiency. Further investigation using multi-omics approaches are in progress to determine the mechanisms by which NCOA4 depletion leads to cell death when extracellular iron supply is limited. Funding Sources Supported by the NIFA, USDA, Hatch project under MIN-18–118 and intramural support to M-S.R.

TAMI-57. INDUCTION OF FERROPTOSIS PROMOTES IMMUNOGENIC CELL DEATH AND ACTIVATION OF THE IMMUNE MICROENVIRONMENT IN GLIOMA

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi210-vi210
Author(s):  
Michael Argenziano ◽  
Matei Banu ◽  
Athanassios Dovas ◽  
Wenting Zhao ◽  
Julia Furnari ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Cell Death ◽  
Antigen Presentation ◽  
Tumor Immunity ◽  
Glioma Cell ◽  
Immunofluorescent Staining ◽  
Immunogenic Cell Death ◽  
Immune Microenvironment ◽  
In Vitro Effects

Abstract Gliomas are immune cold tumors. Effective therapeutic strategies capable of inducing an immune response are lacking.Here we present evidence that ferroptosis, a form of iron-mediated lipid peroxidation-based cell death, may promote anti-tumor immunity via stimulation of phagocytosis and pro-inflammatory activities in microglia. While ferroptosis has shown promise in induction of glioma cell death, the immunogenic and microenvironmental effects of glioma ferroptosis are poorly understood. First, we tested the in vitro effects of the glutathione peroxidase 4 (GPX4) inhibitor RSL3, a ferroptosis inducer, on murine glioma cell lines. Using flow cytometry, we discovered that RSL3 treatment led to membrane translocation of the pro-phagocytic antigen calreticulin, known hallmark of immunogenic cell death, by an average log2-fold-change of 2.53 (p= 0.03) compared to DMSO-treated controls. This effect correlated with lipid peroxidation, as assessed by BODIPY-C11 staining. To further test the effects of ferroptosis on glioma cell-microglia crosstalk, we prepared acute brain tumor slices from both mouse and human glioma samples, and treated them with RSL3. Quantification of immunofluorescent staining from three independent human slice cultures after RSL3 treatment demonstrated a significant increase in calreticulin abundance as compared to control (p < 0.001). Importantly, this effect was significantly diminished with addition of ferrostatin, an inhibitor of ferroptosis, demonstrating that ferroptosis induction was directly responsible for calreticulin translocation. Single-cell RNAseq on mouse and human acute glioma slice cultures treated with RSL3 demonstrated significant overexpression of calreticulin in the tumor population, and positive enrichment of interferon signaling, antigen presentation, and phagocytosis ontologies in both tumor and myeloid compartments. These findings suggest that ferroptosis-induced translocation of calreticulin on the surface of glioma cells promotes activation of the local immune microenvironment by increasing tumor antigen presentation and pro-inflammatory cytokine release by tumor-associated microglia. Thus, ferroptosis-inducing drugs may promote anti-tumor immunity through the activation of immunogenic cell death signals.

Efficacy of Alpha-Lipoic Acid in Iron-Induced Oxidative Stress.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3599-3599
Author(s):  
Ashutosh Lal ◽  
Jung H. Suh ◽  
Bruce N. Ames
Keyword(s):  
Oxidative Stress ◽  
Iron Overload ◽  
Lipoic Acid ◽  
Iron Content ◽  
Redox State ◽  
Ammonium Citrate ◽  
Ferric Ammonium Citrate ◽  
Dependent Manner ◽  
Intracellular Iron ◽  
Dose Dependent

Abstract Antioxidant supplementation could reduce organ damage and premature death in patients with iron overload. We evaluated the improvement in iron-induced oxidative stress with α-Lipoic acid (LA), a multifunctional thiol antioxidant and inducer of phase II enzymes, and compared it with benzylhydroxylamine (BH), a reducing agent that protects against free radicals. Human fibroblasts (IMR-90) were grown in regular cell culture medium or with ferric ammonium citrate (FAC). The intracellular iron content was measured by inductively-coupled plasma spectrometry (ICP). Either LA or BH was added on day 2, and cells grown to confluence. Oxidative stress was assessed by dichlorodihydrofluorescein diacetate fluorescence (DCF-fl). GSH:GSSG was measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The total intracellular iron content (mean±SEM, ng per 5x10^6 cells) increased from 66.95±6.51 in control cells to 262.26±4.69 after exposure to 25 μM FAC, and 556.26±21.15 after exposure to 125 μM FAC. The increase in iron-content of the IMR-90 fibroblasts caused significant rise in oxidant appearance at the highest concentration, as evidenced by 27.8±2.6% increase in steady-state DCF-fl compared with control cells (p <.001). Addition of increasing concentrations of LA attenuated iron-mediated rise in DCF-fl in dose dependent manner. LA at concentrations above 25 μM completely abolished the iron-dependent rise in DCF-fl (91.9±2.3% for FAC+LA, p <.001). BH treated cells also exhibited a dose-dependent decrease in DCF-fl, but a much higher concentration (>200 μM) was needed for complete attenuation of iron-induced rise in DCF-fl (108.3±3.4%, p <.001). These results suggest that LA is more effective than BH in ameliorating iron-mediated increase in oxidative stress. To further understand the basis for the efficacy of LA, intracellular GSH and GSSG were measured. Results show that LA improved the total GSH levels and its redox state in a dose dependent manner. LA and BH attenuated iron-mediated oxidant production, with LA exhibiting greater efficacy. This effect may be due to potent ability of LA to improve intracellular GSH levels and redox state. Our results suggest LA may be useful in reducing complications from iron overload.

scholarly journals Hepatitis C Virus Infection Induces Apoptosis through a Bax-Triggered, Mitochondrion-Mediated, Caspase 3-Dependent Pathway

2008 ◽  
Vol 82 (21) ◽  
pp. 10375-10385 ◽  
Author(s):  
Lin Deng ◽  
Tetsuya Adachi ◽  
Kikumi Kitayama ◽  
Yasuaki Bungyoku ◽  
Sohei Kitazawa ◽  
...  
Keyword(s):  
Cell Death ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Er Stress ◽  
Caspase 3 ◽  
Nuclear Translocation ◽  
Experimental System ◽  
Hcv Infection ◽  
Hcv Rna ◽  
Dependent Pathway

ABSTRACT We previously reported that cells harboring the hepatitis C virus (HCV) RNA replicon as well as those expressing HCV NS3/4A exhibited increased sensitivity to suboptimal doses of apoptotic stimuli to undergo mitochondrion-mediated apoptosis (Y. Nomura-Takigawa, et al., J. Gen. Virol. 87:1935-1945, 2006). Little is known, however, about whether or not HCV infection induces apoptosis of the virus-infected cells. In this study, by using the chimeric J6/JFH1 strain of HCV genotype 2a, we demonstrated that HCV infection induced cell death in Huh7.5 cells. The cell death was associated with activation of caspase 3, nuclear translocation of activated caspase 3, and cleavage of DNA repair enzyme poly(ADP-ribose) polymerase, which is known to be an important substrate for activated caspase 3. These results suggest that HCV-induced cell death is, in fact, apoptosis. Moreover, HCV infection activated Bax, a proapoptotic member of the Bcl-2 family, as revealed by its conformational change and its increased accumulation on mitochondrial membranes. Concomitantly, HCV infection induced disruption of mitochondrial transmembrane potential, followed by mitochondrial swelling and release of cytochrome c from mitochondria. HCV infection also caused oxidative stress via increased production of mitochondrial superoxide. On the other hand, HCV infection did not mediate increased expression of glucose-regulated protein 78 (GRP78) or GRP94, which are known as endoplasmic reticulum (ER) stress-induced proteins; this result suggests that ER stress is not primarily involved in HCV-induced apoptosis in our experimental system. Taken together, our present results suggest that HCV infection induces apoptosis of the host cell through a Bax-triggered, mitochondrion-mediated, caspase 3-dependent pathway(s).

scholarly journals Silibinin induces apoptosis via calpain-dependent AIF nuclear translocation in U87MG human glioma cell death

2011 ◽  
Vol 30 (1) ◽  
pp. 44 ◽  
Author(s):  
Ji C Jeong ◽  
Won Y Shin ◽  
Thae H Kim ◽  
Chae H Kwon ◽  
Jae H Kim ◽  
...  
Keyword(s):  
Cell Death ◽  
Glioma Cell ◽  
Nuclear Translocation ◽  
Human Glioma ◽  
Human Glioma Cell

scholarly journals Regulation of Endothelium-Reticulum-Stress-Mediated Apoptotic Cell Death by a Polymethoxylated Flavone, Nobiletin, Through the Inhibition of Nuclear Translocation of Glyceraldehyde 3-Phosphate Dehydrogenase in Retinal Müller Cells

Cells ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 669
Author(s):  
Yoshiki Miyata ◽  
Kazuya Matsumoto ◽  
Shuichi Kusano ◽  
Yoshio Kusakabe ◽  
Yoshiya Katsura ◽  
...  
Keyword(s):  
Cell Death ◽  
Er Stress ◽  
Nuclear Translocation ◽  
Apoptotic Cell ◽  
Protective Action ◽  
Müller Cells ◽  
Müller Cell ◽  
Nuclear Accumulation ◽  
Muller Cells ◽  
Muller Cell

In the early stages of diabetic retinopathy (DR), subtle biochemical and functional alterations occur in Müller cells, which are one of the components of the blood–retinal barrier (BRB). Müller cells are the principal glia of the retina and have shown a strong involvement in the maintenance of homeostasis and the development of retinal tissue. Their functional abnormalities and eventual loss have been correlated with a decrease in the tight junctions between endothelial cells and a consequent breakdown of the BRB, leading to the development of DR. We demonstrated that the endothelium reticulum (ER) triggers Müller cell death and that nuclear accumulation of glyceraldehyde 3-phosphate dehydrogenase is closely associated with ER-induced Müller cell death. In addition, induction of ER stress in Müller cells increased vascular endothelial growth factor expression but decreased pigment-epithelium-derived factor (PEDF) expression in Müller cells. We found that nobiletin, a polymethoxylated flavone from citrus explants, exerts protective action against ER-stress-induced Müller cell death. In addition, nobiletin was found to augment PEDF expression in Müller cells, which may lead to the protection of BRB integrity. These results suggest that nobiletin can be an attractive candidate for the protection of the BRB from breakdown in DR.

scholarly journals Ferroptosis: the potential value target in atherosclerosis

2021 ◽  
Vol 12 (8) ◽  
Author(s):  
Siyu Ouyang ◽  
Jia You ◽  
Chenxi Zhi ◽  
Pin Li ◽  
Xiaoyan Lin ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Cell Death ◽  
Theoretical Basis ◽  
Vascular Endothelial Cells ◽  
Necrotic Core ◽  
Vascular Endothelial ◽  
Intracellular Iron ◽  
Future Studies ◽  
Dependent Form ◽  
Pathologic Processes

AbstractIn advanced atherosclerosis (AS), defective function-induced cell death leads to the formation of the characteristic necrotic core and vulnerable plaque. The forms and mechanisms of cell death in AS have recently been elucidated. Among them, ferroptosis, an iron-dependent form of necrosis that is characterized by oxidative damage to phospholipids, promotes AS by accelerating endothelial dysfunction in lipid peroxidation. Moreover, disordered intracellular iron causes damage to macrophages, vascular smooth muscle cells (VSMCs), vascular endothelial cells (VECs), and affects many risk factors or pathologic processes of AS such as disturbances in lipid peroxidation, oxidative stress, inflammation, and dyslipidemia. However, the mechanisms through which ferroptosis initiates the development and progression of AS have not been established. This review explains the possible correlations between AS and ferroptosis, and provides a reliable theoretical basis for future studies on its mechanism.

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