Effect of ClAlPcS2 photodynamic and sonodynamic therapy on hela cells

Photodynamic therapy (PDT) uses photosensitive substance to provoke a cytotoxic reaction causing a cell damage or cell death. The substances, photosensitizers, are usually derivates of porphyrine or phtalocyanine. Photosensitizers must be activated by light in order to produce reactive oxygen species, mainly singlet oxygen. Sonodynamic therapy (SDT) utilizes ultrasound to enhance a cytotoxic effects of compounds called sonosensitizers. In this study we investigated photodynamic and sonodynamic effect of chloraluminium phtalocyanine disulfonate (ClAlPcS(2)) on HeLa cells. DNA damage, cell viability and reactive oxygen species (ROS) production were assessed to find whether the combination of PDT and SDT inflicts HeLa cells more than PDT alone. We found that the combined therapy increases DNA fragmentation, enhances ROS production and decreases cell survival. Our results indicate that ClAlPcS(2) can act as a sonosentitiser and combined with PDT causes more irreversible changes to the cells resulting in cell death than PDT alone.

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The Characterization of TA-289, a Novel Antifungal from Fusarium sp.

10.26686/wgtn.16992670 ◽

2021 ◽

Author(s):

◽

Natelle C H Quek

Keyword(s):

Oxidative Stress ◽

Reactive Oxygen Species ◽

Cell Cycle ◽

Cell Death ◽

Reactive Oxygen ◽

Chemical Diversity ◽

Mitochondrial Morphology ◽

Ros Production ◽

Oxygen Species ◽

Wild Type

<p>Natural products offer vast structural and chemical diversity highly sought after in drug discovery research. Saccharomyces cerevisiae makes an ideal model eukaryotic organism for drug mode-of-action studies owing to ease of growth, sophistication of genetic tools and overall homology to higher eukaryotes. Equisetin and a closely related novel natural product, TA-289, are cytotoxic to fermenting yeast, but seemingly less so when yeast actively respire. Cell cycle analyses by flow cytometry revealed a cell cycle block at S-G2/M phase caused by TA-289; previously described oxidative stress-inducing compounds causing cell cycle delay led to further investigation in the involvement of equisetin and TA-289 in mitochondrial-mediated generation of reactive oxygen species. Chemical genomic profiling involving genome-wide scans of yeast deletion mutant strains for TA-289 sensitivity revealed sensitization of genes involved in the mitochondria, DNA damage repair and oxidative stress responses, consistent with a possible mechanism-of-action at the mitochondrion. Flow cytometric detection of reactive oxygen species (ROS) generation caused by TA-289 suggests that the compound may induce cell death via ROS production. The generation of a mutant strain resistant to TA-289 also displayed resistance to a known oxidant, H2O2, at concentrations that were cytotoxic to wild-type cells. The resistant mutant displayed a higher basal level of ROS production compared to the wild-type parent, indicating that the resistance mutation led to an up-regulation of antioxidant capacity which provides cell survival in the presence of TA-289. Yeast mitochondrial morphology was visualized by confocal light microscopy, where it was observed that cells treated with TA-289 displayed abnormal mitochondria phenotypes, further indicating that the compound is acting primarily at the mitochondrion. Similar effects observed with equisetin treatment suggest that both compounds share the same mechanism, eliciting cell death via ROS production in the mitochondrial respiratory chain.</p>

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Survival Signaling by C-RAF: Mitochondrial Reactive Oxygen Species and Ca2+ Are Critical Targets

Molecular and Cellular Biology ◽

10.1128/mcb.00683-07 ◽

2008 ◽

Vol 28 (7) ◽

pp. 2304-2313 ◽

Cited By ~ 31

Author(s):

Andrey V. Kuznetsov ◽

Julija Smigelskaite ◽

Christine Doblander ◽

Manickam Janakiraman ◽

Martin Hermann ◽

...

Keyword(s):

Oxidative Stress ◽

Reactive Oxygen Species ◽

Cell Death ◽

Reactive Oxygen ◽

Ros Production ◽

Oxygen Species ◽

Mitochondrial Reactive Oxygen Species ◽

Mitochondrial Ros ◽

Survival Signaling ◽

First Time

ABSTRACT Survival signaling by RAF occurs through largely unknown mechanisms. Here we provide evidence for the first time that RAF controls cell survival by maintaining permissive levels of mitochondrial reactive oxygen species (ROS) and Ca2+. Interleukin-3 (IL-3) withdrawal from 32D cells resulted in ROS production, which was suppressed by activated C-RAF. Oncogenic C-RAF decreased the percentage of apoptotic cells following treatment with staurosporine or the oxidative stress-inducing agent tert-butyl hydroperoxide. However, it was also the case that in parental 32D cells growing in the presence of IL-3, inhibition of RAF signaling resulted in elevated mitochondrial ROS and Ca2+ levels. Cell death is preceded by a ROS-dependent increase in mitochondrial Ca2+, which was absent from cells expressing transforming C-RAF. Prevention of mitochondrial Ca2+ overload after IL-3 deprivation increased cell viability. MEK was essential for the mitochondrial effects of RAF. In summary, our data show that survival control by C-RAF involves controlling ROS production, which otherwise perturbs mitochondrial Ca2+ homeostasis.

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

Evidence-based Complementary and Alternative Medicine ◽

10.1093/ecam/nep082 ◽

2011 ◽

Vol 2011 ◽

pp. 1-12 ◽

Cited By ~ 21

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.

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Transient Increase in Cellular Dehydrogenase Activity After Cadmium Treatment Precedes Enhanced Production of Reactive Oxygen Species in Human Proximal Tubular Kidney Cells

Physiological Research ◽

10.33549/physiolres.934121 ◽

2019 ◽

pp. 481-490 ◽

Cited By ~ 1

Author(s):

J. HANDL ◽

J. ČAPEK ◽

P. MAJTNEROVÁ ◽

F. PETIRA ◽

M. HAUSCHKE ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Cell Death ◽

Dehydrogenase Activity ◽

Reactive Oxygen ◽

Transient Increase ◽

Kidney Cells ◽

Ros Production ◽

Oxygen Species ◽

Cadmium Treatment ◽

Jnk Activation

Cadmium is a heavy metal causing toxicity especially in kidney cells. The toxicity is linked also with enhanced oxidative stress leading to cell death. On the other hand, our recent experiments have shown that an increase of total intracellular dehydrogenases activity can also occur in kidney cells before declining until cell death. The aim of the present study, therefore, was to evaluate this transient enhancement in cell viability after cadmium treatment. The human kidney HK-2 cell line was treated with CdCl2 at concentrations 0-200 µM for 2-24 h and intracellular dehydrogenase activity was tested. In addition, we measured reactive oxygen species (ROS) production, glutathione levels, mitochondrial membrane potential, and C-Jun-N-terminal kinase (JNK) activation. We found that significantly increased dehydrogenase activity could occur in cells treated with 25, 100, and 200 µM CdCl2. Moreover, the results showed an increase in ROS production linked with JNK activation following the enhancement of dehydrogenase activity. Other tests detected no relationship with the increased in intracellular dehydrogenase activity. Hence, the transient increase in dehydrogenase activity in HK-2 cells preceded the enhancement of ROS production and our finding provides new evidence in cadmium kidney toxicity.

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Reoxygenation after hypoxia and glucose depletion causes reactive oxygen species production by mitochondria in HUVEC

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.00048.2004 ◽

2004 ◽

Vol 287 (5) ◽

pp. R1037-R1043 ◽

Cited By ~ 52

Author(s):

S. Therade-Matharan ◽

E. Laemmel ◽

J. Duranteau ◽

E. Vicaut

Keyword(s):

Reactive Oxygen Species ◽

Cell Death ◽

Respiratory Chain ◽

Reactive Oxygen ◽

Chain Complex ◽

Ros Production ◽

Oxygen Species ◽

Respiratory Chain Complex ◽

Type Iii ◽

Glucose Depletion

In hemorrhagic shock, local hypoxia is present and followed by reoxygenation during the therapeutic process. In endothelium, reactive oxygen species (ROS) have been identified as a cause of inflammatory reactions and tissular lesions in ischemic territory during reoxygenation. This study was designed to identify the enzymatic mechanisms of ROS formation during reoxygenation after hypoxia. Because severe shock, in vivo, can affect both O2 and nutriments, we combined hypoxia at a level close to that found in terminal vessels during shock, with glucose depletion, which induces a relevant additional stress. Human umbilical vein endothelial cells (HUVEC) underwent 2 h of hypoxia (Po2 ∼20 mmHg) without glucose and 1 h of reoxygenation (Po2 ∼120 mmHg) with glucose. ROS production was measured by the fluorescent marker 2′,7′-dichlorodihydrofluorescein diacetate, and cell death by propidium iodide. After 1 h of reoxygenation, fluorescence had risen by 143 ± 17%. Cell death was equal to 8.6 ± 2.4%. Antimycin A and stigmatellin, which inhibits the type III mitochondrial respiratory chain complex, reduced ROS production to values of 61 ± 10 and 59 ± 7%, respectively, but inhibitors of other chain complexes did not affect it. In addition, the increase in fluorescence was not affected by inhibition of NADPH oxidase, xanthine oxidase, NOS, cyclooxygenase, cytochrome P-450 monooxygenase, or monoamine oxidase. We did not observe any increase in cell death. These results show that, in HUVEC, mitochondria are responsible for ROS production after hypoxia and reoxygenation and suggest that a ROS release site is activated in the cytochrome b of the type III respiratory chain complex.

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The Regulation of Reactive Oxygen Species Production during Programmed Cell Death

The Journal of Cell Biology ◽

10.1083/jcb.141.6.1423 ◽

1998 ◽

Vol 141 (6) ◽

pp. 1423-1432 ◽

Cited By ~ 505

Author(s):

Shirlee Tan ◽

Yutaka Sagara ◽

Yuanbin Liu ◽

Pamela Maher ◽

David Schubert

Keyword(s):

Gene Expression ◽

Reactive Oxygen Species ◽

Cell Death ◽

Programmed Cell Death ◽

Reactive Oxygen ◽

Fold Increase ◽

Glutamate Toxicity ◽

Ros Production ◽

Oxygen Species ◽

Ht22 Cells

Reactive oxygen species (ROS) are thought to be involved in many forms of programmed cell death. The role of ROS in cell death caused by oxidative glutamate toxicity was studied in an immortalized mouse hippocampal cell line (HT22). The causal relationship between ROS production and glutathione (GSH) levels, gene expression, caspase activity, and cytosolic Ca2+ concentration was examined. An initial 5–10-fold increase in ROS after glutamate addition is temporally correlated with GSH depletion. This early increase is followed by an explosive burst of ROS production to 200–400-fold above control values. The source of this burst is the mitochondrial electron transport chain, while only 5–10% of the maximum ROS production is caused by GSH depletion. Macromolecular synthesis inhibitors as well as Ac-YVAD-cmk, an interleukin 1β–converting enzyme protease inhibitor, block the late burst of ROS production and protect HT22 cells from glutamate toxicity when added early in the death program. Inhibition of intracellular Ca2+ cycling and the influx of extracellular Ca2+ also blocks maximum ROS production and protects the cells. The conclusion is that GSH depletion is not sufficient to cause the maximal mitochondrial ROS production, and that there is an early requirement for protease activation, changes in gene expression, and a late requirement for Ca2+ mobilization.

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SWCNT–porphyrin nano-hybrids selectively activated by ultrasound: an interesting model for sonodynamic applications

RSC Advances ◽

10.1039/d0ra03944f ◽

2020 ◽

Vol 10 (37) ◽

pp. 21736-21744 ◽

Cited By ~ 1

Author(s):

Federica Bosca ◽

Ingrid Corazzari ◽

Federica Foglietta ◽

Roberto Canaparo ◽

Gianni Durando ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Cell Death ◽

Reactive Oxygen ◽

Acoustic Cavitation ◽

Oxygen Species ◽

Sonodynamic Therapy

Sonodynamic therapy (SDT) is an innovative anticancer approach, based on the excitation of a given molecule (usually a porphyrin) by inertial acoustic cavitation that leads to cell death via the production of reactive oxygen species (ROS).

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Antioxidants Threaten Multikinase Inhibitor Efficacy against Liver Cancer by Blocking Mitochondrial Reactive Oxygen Species

Antioxidants ◽

10.3390/antiox10091336 ◽

2021 ◽

Vol 10 (9) ◽

pp. 1336

Author(s):

Blanca Cucarull ◽

Anna Tutusaus ◽

Tania Hernáez-Alsina ◽

Pablo García de Frutos ◽

María Reig ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Cell Death ◽

Liver Cancer ◽

Cell Lines ◽

Reactive Oxygen ◽

Chemotherapeutic Agents ◽

Glutathione Depletion ◽

Ros Production ◽

Oxygen Species ◽

Tumor Spheroids

Sorafenib and regorafenib, multikinase inhibitors (MKIs) used as standard chemotherapeutic agents for hepatocellular carcinoma (HCC), generate reactive oxygen species (ROS) during cancer treatment. Antioxidant supplements are becoming popular additions to our diet, particularly glutathione derivatives and mitochondrial-directed compounds. To address their possible interference during HCC chemotherapy, we analyzed the effect of common antioxidants using hepatoma cell lines and tumor spheroids. In liver cancer cell lines, sorafenib and regorafenib induced mitochondrial ROS production and potent cell death after glutathione depletion. In contrast, cabozantinib only exhibited oxidative cell death in specific HCC cell lines. After sorafenib and regorafenib administration, antioxidants such as glutathione methyl ester and the superoxide scavenger MnTBAP decreased cell death and ROS production, precluding the MKI activity against hepatoma cells. Interestingly, sorafenib-induced mitochondrial damage caused PINK/Parkin-dependent mitophagy stimulation, altered by increased ROS production. Finally, in sorafenib-treated tumor spheroids, while ROS induction reduced tumor growth, antioxidant treatments favored tumor development. In conclusion, the anti-tumor activity of specific MKIs, such as regorafenib and sorafenib, is altered by the cellular redox status, suggesting that uncontrolled antioxidant intake during HCC treatment should be avoided or only endorsed to diminish chemotherapy-induced side effects, always under medical scrutiny.

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Role of reactive oxygen species in acetylcholine-induced preconditioning in cardiomyocytes

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1999.277.6.h2504 ◽

1999 ◽

Vol 277 (6) ◽

pp. H2504-H2509 ◽

Cited By ~ 34

Author(s):

Zhenhai Yao ◽

Jiankun Tong ◽

Xiaohui Tan ◽

Changqing Li ◽

Zuohui Shao ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Cell Death ◽

Ventricular Myocytes ◽

Reactive Oxygen ◽

Ros Production ◽

Oxygen Species ◽

Intracellular Signals ◽

Dichlorofluorescin Diacetate ◽

Flow Through

We examined the ability of ACh to mimic ischemic preconditioning in cardiomyocytes and the role of ATP-sensitive potassium (KATP) channels and mitochondrial reactive oxygen species (ROS) in mediating this effect. Chick embryonic ventricular myocytes were studied in a flow-through chamber while flow rate, pH,[Formula: see text], and[Formula: see text] were controlled. Cell viability was quantified with propidium iodide (5 μM), and production of ROS was measured using 2′,7′-dichlorofluorescin diacetate. Data were expressed as means ± SE. Preconditioning with 10 min of ischemia followed by 10 min of reoxygenation or 10 min of ACh (1 mM) followed by a drug-free period before 1 h of ischemia and 3 h of reoxygenation reduced cell death to the same extent [preconditioning 19 ± 2% ( n = 6, P < 0.05) ACh 21 ± 5% ( n = 6, P < 0.05) vs controls 42 ± 5% ( n = 9)]. Like preconditioning, ACh increased ROS production threefold before ischemia [0.60 ± 0.16 ( n = 7, P< 0.05) vs. controls, 0.16 ± 0.03 ( n = 6); arbitrary units]. Protection and increased ROS production during ACh preconditioning were abolished with 5-hydroxydecanoate (5-HD, 100 μM), a selective mitochondrial KATP channel antagonist, and the thiol reductant 2-mercaptopropionyl glycine (2-MPG, 1 mM), an antioxidant [cell death: 5-HD+ACh 37 ± 7% ( n = 5), 2-MPG+ACh 47 ± 6% ( n = 6); ROS signals: 5-HD+ACh 0.09 ± 0.03 ( n = 5), 2-MPG+ACh 0.01 ± 0.04 ( n = 4)]. In addition, ACh-induced ROS signaling was blocked by the mitochondrial site III electron transport inhibitor myxothiazol (0.02 ± 0.07, n = 5). These results demonstrate that activation of mitochondrial KATPchannels and increased ROS production from mitochondria are important intracellular signals that participate in ACh-induced preconditioning in cardiomyocytes.

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Chondrocyte cell death mediated by reactive oxygen species-dependent activation of PKC-βI

AJP Cell Physiology ◽

10.1152/ajpcell.00214.2005 ◽

2006 ◽

Vol 290 (3) ◽

pp. C802-C811 ◽

Cited By ~ 30

Author(s):

Marcello DelCarlo ◽

Richard F. Loeser

Keyword(s):

Reactive Oxygen Species ◽

Cell Death ◽

Cell Survival ◽

Time Course ◽

Reactive Oxygen ◽

Specific Protein ◽

Ros Production ◽

Oxygen Species ◽

Secondary Messenger ◽

Promote Cell Survival

Signals generated by the extracellular matrix (ECM) promote cell survival. We have shown that chondrocytes detached from their native ECM and plated without serum at low density on poly-l-lysine undergo significant cell death that is associated with the production of reactive oxygen species (ROS). No cell death or ROS production was observed when cells were plated on fibronectin under the same conditions. Cell death on poly-l-lysine could be completely inhibited with the addition of either antioxidants or inhibitors of specific protein kinase C (PKC) isoforms including PKC-βI. PKC-βI was noted to translocate from the cytosol to the particulate membrane after plating on poly-l-lysine, and this translocation was inhibited by the addition of an antioxidant. Time-course analyses implicated endogenous ROS production as a secondary messenger leading to PKC-βI activation and subsequent chondrocyte cell death. Cell survival on poly-l-lysine was significantly improved in the presence of oligomycin or DIDS, suggesting that ROS production occurred via complex V of the electron transport chain of the mitochondria and that ROS were released to the cytosol via voltage-dependent anion channels. Together, these results represent a novel mechanism by which ROS can initiate cell death through the activation of PKC-βI.

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