Hydrogen Peroxide Toxicity Induces Ras Signaling in Human Neuroblastoma SH-SY5Y Cultured Cells

It has been reported that overproduction of reactive oxygen species occurs after brain injury and mediates neuronal cells degeneration. In the present study, we examined the role of Ras signaling on hydrogen peroxide-induced neuronal cells degeneration in dopaminergic neuroblastoma SH-SY5Y cells. Hydrogen peroxide significantly reduced cell viability in SH-SY5Y cultured cells. An inhibitor of the enzyme that catalyzes the farnesylation of Ras proteins, FTI-277, and a competitive inhibitor of GTP-binding proteins, GDP-beta-S significantly decreased hydrogen peroxide-induced reduction in cell viability in SH-SY5Y cultured cells. The results of this study might indicate that a Ras-dependent signaling pathway plays a role in hydrogen peroxide-induced toxicity in neuronal cells.

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ROLE OF PREGNAN-X-RECEPTOR IN CELL RESISTANCE TO NITROSATIVE AND OXIDATIVE STRESS

10.47501/978-5-6044060-1-4.47 ◽

2021 ◽

Author(s):

Yulia Abalenikhina ◽

◽

Elena A. Sudakova ◽

Pelageya Erokhina ◽

Aleksey Shchulkin ◽

...

Keyword(s):

Oxidative Stress ◽

Hydrogen Peroxide ◽

Cell Viability ◽

Nitrosative Stress ◽

Pregnane X Receptor ◽

Cell Resistance ◽

Oxidative And Nitrosative Stress ◽

High Concentrations ◽

And Oxidative Stress

The article discusses the new role of pregnane X receptor (PXR) under conditions of oxidative and nitrosative stress. The results showed that the effect of hydrogen peroxide and S-nitrosoglu-tathione in high concentrations on Caco-2 cells leads to a decrease in cell viability, which is accompanied by an increase in the amount of PXR. These changes are offset by the addition of ketoconazole (inhibitor of PXR) to the medium.

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Stimulatory Heterotrimeric GTP-binding Protein Inhibits Hydrogen Peroxide-induced Apoptosis by Repressing BAK Induction in SH-SY5Y Human Neuroblastoma Cells

Journal of Biological Chemistry ◽

10.1074/jbc.m702344200 ◽

2007 ◽

Vol 283 (3) ◽

pp. 1350-1361 ◽

Cited By ~ 24

Author(s):

So-Young Kim ◽

MiRan Seo ◽

Yeni Kim ◽

Yun-Il Lee ◽

Jung-Min Oh ◽

...

Keyword(s):

Hydrogen Peroxide ◽

Cancer Cells ◽

Neuroblastoma Cells ◽

Camp Signaling ◽

Human Neuroblastoma Cells ◽

Human Neuroblastoma ◽

Gtp Binding ◽

Camp Signaling Pathway ◽

Induced Apoptosis

Heterotrimeric stimulatory GTP-binding protein (Gs) stimulates adenylate cyclases to activate the cAMP signaling pathway. Although the cAMP pathway has been reported to be involved in apoptosis, the role of the Gs-cAMP signaling pathway during reactive oxygen species (ROS)-mediated apoptosis, which is involved in the resistance of cancer cells to chemotherapy and radiation, is not clearly understood. Thus, in this study we aimed to investigate the role of the α subunit of Gs (Gαs) in the ROS-induced apoptosis of cancer cells. The stable expression of constitutively active Gαs (GαsQL) inhibited the hydrogen peroxide-induced apoptosis of SH-SY5Y human neuroblastoma cells and reduced the hydrogen peroxide-induced increase in Bak and the decrease in Bcl-xL protein expression. Exogenous Bak expression abolished these inhibitory effects of GαsQL, but Bak small interfering RNA decreased hydrogen peroxide-induced apoptosis. Gαs repressed hydrogen peroxide-induced Bak expression by inhibiting the transcription of Bak mRNA, which resulted from the inhibition of the hydrogen peroxide-induced activation of transcription factors such as AP1, NF-κB, and NFAT. Moreover, Gαs also inhibited the hydrogen peroxide-induced binding of AP1, NF-κB, and NFAT to the Bak promoter. Furthermore, hydrogen peroxide-induced apoptosis was reduced by treating cells with prostaglandin E2, which activates Gαs, but this was augmented by CCPA, which activates Gαi causing a decrease in cAMP levels. From the results, we conclude that Gαs protects neuroblastoma cells from hydrogen peroxide-induced apoptosis by repressing Bak induction, which is mediated by the inhibition of the hydrogen peroxide-induced activations of AP1, NF-κB, and NFAT through cAMP-PKA-CREB signaling system.

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Protective Role of Catalase in Pseudomonas aeruginosa Biofilm Resistance to Hydrogen Peroxide

Applied and Environmental Microbiology ◽

10.1128/aem.65.10.4594-4600.1999 ◽

1999 ◽

Vol 65 (10) ◽

pp. 4594-4600 ◽

Cited By ~ 148

Author(s):

James G. Elkins ◽

Daniel J. Hassett ◽

Philip S. Stewart ◽

Herbert P. Schweizer ◽

Timothy R. McDermott

Keyword(s):

Hydrogen Peroxide ◽

Pseudomonas Aeruginosa ◽

Single Dose ◽

Cell Viability ◽

Catalase Activity ◽

Type Strain ◽

Wild Type Strain ◽

Wild Type ◽

Planktonic Cells

ABSTRACT The role of the two known catalases in Pseudomonas aeruginosa in protecting planktonic and biofilm cells against hydrogen peroxide (H2O2) was investigated. Planktonic cultures and biofilms formed by the wild-type strain PAO1 and the katA and katB catalase mutants were compared for their susceptibility to H2O2. Over the course of 1 h, wild-type cell viability decreased steadily in planktonic cells exposed to a single dose of 50 mM H2O2, whereas biofilm cell viability remained at approximately 90% when cells were exposed to a flowing stream of 50 mM H2O2. The katB mutant, lacking the H2O2-inducible catalase KatB, was similar to the wild-type strain with respect to H2O2 resistance. The katA mutant possessed undetectable catalase activity. PlanktonickatA mutant cultures were hypersusceptible to a single dose of 50 mM H2O2, while biofilms displayed a 10-fold reduction in the number of culturable cells after a 1-h exposure to 50 mM H2O2. Catalase activity assays, activity stains in nondenaturing polyacrylamide gels, andlacZ reporter genes were used to characterize the oxidative stress responses of planktonic cultures and biofilms. Enzyme assays and catalase activity bands in nondenaturing polyacrylamide gels showed significant KatB catalase induction occurred in biofilms after a 20-min exposure to H2O2, suggesting that biofilms were capable of a rapid adaptive response to the oxidant. Reporter gene data obtained with a katB::lacZtranscriptional reporter strain confirmed katB induction and that the increase in total cellular catalase activity was attributable to KatB. Biofilms upregulated the reporter in the constant presence of 50 mM H2O2, while planktonic cells were overwhelmed by a single 50 mM dose and were unable to make detectable levels of β-galactosidase. The results of this study demonstrated the following: the constitutively expressed KatA catalase is important for resistance of planktonic and biofilm P. aeruginosa to H2O2, particularly at high H2O2 concentrations; KatB is induced in both planktonic and biofilm cells in response to H2O2 insult, but plays a relatively small role in biofilm resistance; and KatB is important to either planktonic cells or biofilm cells for acquired antioxidant resistance when initial levels of H2O2 are sublethal.

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Plasma Membrane Localization Is Required for RasA-Mediated Polarized Morphogenesis and Virulence of Aspergillus fumigatus

Eukaryotic Cell ◽

10.1128/ec.00091-12 ◽

2012 ◽

Vol 11 (8) ◽

pp. 966-977 ◽

Cited By ~ 31

Author(s):

Jarrod R. Fortwendel ◽

Praveen R. Juvvadi ◽

Luise E. Rogg ◽

Yohannes G. Asfaw ◽

Kimberlie A. Burns ◽

...

Keyword(s):

Cell Wall ◽

Plasma Membrane ◽

Aspergillus Fumigatus ◽

Structural Changes ◽

Hyphal Growth ◽

Membrane Association ◽

Ras Signaling ◽

Ras Proteins ◽

Content Type

ABSTRACT Ras is a highly conserved GTPase protein that is essential for proper polarized morphogenesis of filamentous fungi. Localization of Ras proteins to the plasma membrane and endomembranes through posttranslational addition of farnesyl and palmitoyl residues is an important mechanism through which cells provide specificity to Ras signal output. Although the Aspergillus fumigatus RasA protein is known to be a major regulator of growth and development, the membrane distribution of RasA during polarized morphogenesis and the role of properly localized Ras signaling in virulence of a pathogenic mold remain unknown. Here we demonstrate that Aspergillus fumigatus RasA localizes primarily to the plasma membrane of actively growing hyphae. We show that treatment with the palmitoylation inhibitor 2-bromopalmitate disrupts normal RasA plasma membrane association and decreases hyphal growth. Targeted mutations of the highly conserved RasA palmitoylation motif also mislocalized RasA from the plasma membrane and led to severe hyphal abnormalities, cell wall structural changes, and reduced virulence in murine invasive aspergillosis. Finally, we provide evidence that proper RasA localization is independent of the Ras palmitoyltransferase homolog, encoded by erfB , but requires the palmitoyltransferase complex subunit, encoded by erfD . Our results demonstrate that plasma membrane-associated RasA is critical for polarized morphogenesis, cell wall stability, and virulence in A. fumigatus .

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The Role of Wild-Type RAS in Oncogenic RAS Transformation

Genes ◽

10.3390/genes12050662 ◽

2021 ◽

Vol 12 (5) ◽

pp. 662

Author(s):

Erin Sheffels ◽

Robert L. Kortum

Keyword(s):

Family Members ◽

Protein Product ◽

Therapeutic Resistance ◽

Ras Signaling ◽

Ras Proteins ◽

Wild Type ◽

Oncogenic Ras ◽

Downstream Effector ◽

Ras Family

The RAS family of oncogenes (HRAS, NRAS, and KRAS) are among the most frequently mutated protein families in cancers. RAS-mutated tumors were originally thought to proliferate independently of upstream signaling inputs, but we now know that non-mutated wild-type (WT) RAS proteins play an important role in modulating downstream effector signaling and driving therapeutic resistance in RAS-mutated cancers. This modulation is complex as different WT RAS family members have opposing functions. The protein product of the WT RAS allele of the same isoform as mutated RAS is often tumor-suppressive and lost during tumor progression. In contrast, RTK-dependent activation of the WT RAS proteins from the two non-mutated WT RAS family members is tumor-promoting. Further, rebound activation of RTK–WT RAS signaling underlies therapeutic resistance to targeted therapeutics in RAS-mutated cancers. The contributions of WT RAS to proliferation and transformation in RAS-mutated cancer cells places renewed interest in upstream signaling molecules, including the phosphatase/adaptor SHP2 and the RasGEFs SOS1 and SOS2, as potential therapeutic targets in RAS-mutated cancers.

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Role of nitric oxide synthase in release of endothelin from cultured human endometrial cells

Acta Endocrinologica ◽

10.1530/eje.0.1380467 ◽

1998 ◽

pp. 467-471 ◽

Cited By ~ 3

Author(s):

Z Lin ◽

T Kubota ◽

M Masuda ◽

T Aso

Keyword(s):

Nitric Oxide ◽

Nitric Oxide Synthase ◽

Cultured Cells ◽

Close Association ◽

Competitive Inhibitor ◽

Secretory Phase ◽

Endometrial Stromal Cells ◽

Endometrial Cells ◽

Oxide Synthase

The aim of this study was to investigate the influence of the nitric oxide/nitric oxide synthase (NO/NOS) system on the release of endothelin-1 (ET-1) in human endometrial cells. Human endometrial stromal cells in secretory phase were incubated for 72 h in serum-free RPMI 1640 medium in the absence or presence of different concentrations of interleukin-1beta (IL-1beta) and NG-monomethyl-L-arginine (LNMMA), a specific competitive inhibitor of NOS. ET-1 released from the cultured cells into the medium was determined by specific RIA. In all the experiments at various times, IL-1beta significantly increased the release of ET-1. LNMMA significantly attenuated the release of ET-1 when the cells were cultured with both IL-1beta and LNMMA, but LNMMA alone had no effect on ET-1 release. These results suggest that the NO/NOS system in human endometrium is involved in the regulation of ET-1 release via IL-1beta secretion. It can also be inferred that NO and ET-1 control the functions of endometrium in close association with IL-1beta.

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Protective role of Populus tomentiglandulosa against hydrogen peroxide-induced oxidative stress in SH-SY5Y neuronal cells

Journal of Applied Biological Chemistry ◽

10.3839/jabc.2020.047 ◽

2020 ◽

Vol 63 (4) ◽

pp. 357-363

Author(s):

Yu Ri Kwon ◽

Ji Hyun Kim ◽

Sanghyun Lee ◽

Eun Ju Cho ◽

Hyun Young Kim

Keyword(s):

Oxidative Stress ◽

Hydrogen Peroxide ◽

Neuronal Cells ◽

Protective Role

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Mutations in LZTR1 drive human disease by dysregulating RAS ubiquitination

Science ◽

10.1126/science.aap7607 ◽

2018 ◽

Vol 362 (6419) ◽

pp. 1177-1182 ◽

Cited By ~ 52

Author(s):

M. Steklov ◽

S. Pandolfi ◽

M. F. Baietti ◽

A. Batiuk ◽

P. Carai ◽

...

Keyword(s):

Ubiquitin Ligase ◽

Human Disease ◽

Mammalian Cells ◽

Leucine Zipper ◽

Ras Signaling ◽

Ras Proteins ◽

Ubiquitin Ligase Complex ◽

Cullin 3 ◽

Guanosine Triphosphatase

The leucine zipper–like transcriptional regulator 1 (LZTR1) protein, an adaptor for cullin 3 (CUL3) ubiquitin ligase complex, is implicated in human disease, yet its mechanism of action remains unknown. We found that Lztr1 haploinsufficiency in mice recapitulates Noonan syndrome phenotypes, whereas LZTR1 loss in Schwann cells drives dedifferentiation and proliferation. By trapping LZTR1 complexes from intact mammalian cells, we identified the guanosine triphosphatase RAS as a substrate for the LZTR1-CUL3 complex. Ubiquitome analysis showed that loss of Lztr1 abrogated Ras ubiquitination at lysine-170. LZTR1-mediated ubiquitination inhibited RAS signaling by attenuating its association with the membrane. Disease-associated LZTR1 mutations disrupted either LZTR1-CUL3 complex formation or its interaction with RAS proteins. RAS regulation by LZTR1-mediated ubiquitination provides an explanation for the role of LZTR1 in human disease.

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Role of extracellular Hydrogen peroxide in regulation of iron homeostasis genes in neuronal cells: Implication in iron accumulation

Free Radical Biology and Medicine ◽

10.1016/j.freeradbiomed.2015.05.025 ◽

2015 ◽

Vol 86 ◽

pp. 78-89 ◽

Cited By ~ 16

Author(s):

Som Dev ◽

Sanju Kumari ◽

Neena Singh ◽

Saswat Kumar Bal ◽

Pankaj Seth ◽

...

Keyword(s):

Hydrogen Peroxide ◽

Iron Homeostasis ◽

Neuronal Cells ◽

Iron Accumulation

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Cytoprotective Effect of Tocotrienol-Rich Fraction and α-Tocopherol Vitamin E Isoforms Against Glutamate-Induced Cell Death in Neuronal Cells

International Journal for Vitamin and Nutrition Research ◽

10.1024/0300-9831/a000201 ◽

2014 ◽

Vol 84 (3-4) ◽

pp. 0140-0151 ◽

Cited By ~ 11

Author(s):

Thilaga Rati Selvaraju ◽

Huzwah Khaza’ai ◽

Sharmili Vidyadaran ◽

Mohd Sokhini Abd Mutalib ◽

Vasudevan Ramachandran ◽

...

Keyword(s):

Vitamin E ◽

Cell Viability ◽

Neuronal Cells ◽

Positive Control ◽

Post Treatment ◽

Mitochondrial Activity ◽

Before And After ◽

Pre Treatment ◽

Tocotrienol Rich Fraction ◽

Major Mediator

Glutamate is the major mediator of excitatory signals in the mammalian central nervous system. Extreme amounts of glutamate in the extracellular spaces can lead to numerous neurodegenerative diseases. We aimed to clarify the potential of the following vitamin E isomers, tocotrienol-rich fraction (TRF) and α-tocopherol (α-TCP), as potent neuroprotective agents against glutamate-induced injury in neuronal SK-N-SH cells. Cells were treated before and after glutamate injury (pre- and post-treatment, respectively) with 100 - 300 ng/ml TRF/α-TCP. Exposure to 120 mM glutamate significantly reduced cell viability to 76 % and 79 % in the pre- and post-treatment studies, respectively; however, pre- and post-treatment with TRF/α-TCP attenuated the cytotoxic effect of glutamate. Compared to the positive control (glutamate-injured cells not treated with TRF/α-TCP), pre-treatment with 100, 200, and 300 ng/ml TRF significantly improved cell viability following glutamate injury to 95.2 %, 95.0 %, and 95.6 %, respectively (p < 0.05).The isomers not only conferred neuroprotection by enhancing mitochondrial activity and depleting free radical production, but also increased cell viability and recovery upon glutamate insult. Our results suggest that vitamin E has potent antioxidant potential for protecting against glutamate injury and recovering glutamate-injured neuronal cells. Our findings also indicate that both TRF and α-TCP could play key roles as anti-apoptotic agents with neuroprotective properties.

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