Glutamate Toxicity on a PC12 Cell Line Involves Glutathione (GSH) Depletion and Oxidative Stress

Sulfur mustard (SM) is a chemical warfare agent that can damage DNA via alkylation and oxidative stress. Because of its genotoxicity, SM is cancerogenic and the progenitor of many chemotherapeutics. Previously, we developed an SM-resistant cell line via chronic exposure of the popular keratinocyte cell line HaCaT to increasing doses of SM over a period of 40 months. In this study, we compared the genomic landscape of the SM-resistant cell line HaCaT/SM to its sensitive parental line HaCaT in order to gain insights into genetic changes associated with continuous alkylation and oxidative stress. We established chromosome numbers by cytogenetics, analyzed DNA copy number changes by means of array Comparative Genomic Hybridization (array CGH), employed the genome-wide chromosome conformation capture technique Hi-C to detect chromosomal translocations, and derived mutational signatures by whole-genome sequencing. We observed that chronic SM exposure eliminated the initially prevailing hypotetraploid cell population in favor of a hyperdiploid one, which contrasts with previous observations that link polyploidization to increased tolerance and adaptability toward genotoxic stress. Furthermore, we observed an accumulation of chromosomal translocations, frequently flanked by DNA copy number changes, which indicates a high rate of DNA double-strand breaks and their misrepair. HaCaT/SM-specific single-nucleotide variants showed enrichment of C > A and T > A transversions and a lower rate of deaminated cytosines in the CpG dinucleotide context. Given the frequent use of HaCaT in toxicology, this study provides a valuable data source with respect to the original genotype of HaCaT and the mutational signatures associated with chronic alkylation and oxidative stress.

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Biotransformation and oxidative stress responses in rat hepatic cell-line (H4IIE) exposed to organophosphate esters (OPEs)

Toxicology and Applied Pharmacology ◽

10.1016/j.taap.2019.04.004 ◽

2019 ◽

Vol 371 ◽

pp. 84-94 ◽

Cited By ~ 11

Author(s):

Elvira Mennillo ◽

Francesca Cappelli ◽

Augustine Arukwe

Keyword(s):

Oxidative Stress ◽

Cell Line ◽

Stress Responses ◽

Hepatic Cell ◽

Organophosphate Esters ◽

Hepatic Cell Line ◽

Oxidative Stress Responses ◽

And Oxidative Stress

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Suppression of proliferation and oxidative stress by extracts of Ganoderma lucidum in the ovarian cancer cell line OVCAR-3

International Journal of Molecular Medicine ◽

10.3892/ijmm.2011.788 ◽

2011 ◽

Cited By ~ 7

Author(s):

Joseph Wu

Keyword(s):

Oxidative Stress ◽

Ovarian Cancer ◽

Cell Line ◽

Cancer Cell ◽

Ganoderma Lucidum ◽

Ovarian Cancer Cell ◽

Ovarian Cancer Cell Line ◽

Cancer Cell Line ◽

And Oxidative Stress

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Establishment and Characterization of Methylmercury-Resistant PC12 Cell Line

Environmental Health Perspectives ◽

10.2307/3431810 ◽

1994 ◽

Vol 102 ◽

pp. 313 ◽

Cited By ~ 1

Author(s):

Kyoko Miura ◽

Thomas W. Clarkson ◽

Kazumasa Ikeda ◽

Akira Naganuma ◽

Nobumasa Imura

Keyword(s):

Cell Line ◽

Pc12 Cell ◽

Pc12 Cell Line

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Synaptotagmin VII Is Targeted to Dense-core Vesicles and Regulates Their Ca2+-dependent Exocytosis in PC12 Cells

Journal of Biological Chemistry ◽

10.1074/jbc.m409241200 ◽

2004 ◽

Vol 279 (50) ◽

pp. 52677-52684 ◽

Cited By ~ 58

Author(s):

Mitsunori Fukuda ◽

Eiko Kanno ◽

Megumi Satoh ◽

Chika Saegusa ◽

Akitsugu Yamamoto

Keyword(s):

Plasma Membrane ◽

Neuropeptide Y ◽

Cell Line ◽

Pc12 Cells ◽

Pc12 Cell ◽

Small Interfering Rna ◽

Dense Core ◽

Dense Core Vesicles ◽

Pc12 Cell Line ◽

Interfering Rna

It has recently been proposed that synaptotagmin (Syt) VII functions as a plasma membrane Ca2+sensor for dense-core vesicle exocytosis in PC12 cells based on the results of transient overexpression studies using green fluorescent protein (GFP)-tagged Syt VII; however, the precise subcellular localization of Syt VII is still a matter of controversy (plasma membraneversussecretory granules). In this study we established a PC12 cell line “stably expressing” the Syt VII-GFP molecule and demonstrated by immunocytochemical and immunoelectron microscopic analyses that the Syt VII-GFP protein is localized on dense-core vesicles as well as in other intracellular membranous structures, such as thetrans-Golgi network and lysosomes. Syt VII-GFP forms a complex with endogenous Syts I and IX, but not with Syt IV, and it colocalize well with Syts I and IX in the cellular processes (where dense-core vesicles are accumulated) in the PC12 cell line. We further demonstrated by an N-terminal antibody-uptake experiment that Syt VII-GFP-containing dense-core vesicles undergo Ca2+-dependent exocytosis, the same as endogenous Syt IX-containing vesicles. Moreover, silencing of Syt VII-GFP with specific small interfering RNA dramatically reduced high KCl-dependent neuropeptide Y secretion from the stable PC12 cell line (∼60% of the control cells), whereas the same small interfering RNA had little effect on neuropeptide Y secretion from the wild-type PC12 cells (∼85–90% of the control cells), indicating that the level of endogenous expression of Syt VII molecules must be low. Our results indicate that the targeting of Syt VII-GFP molecules to specific membrane compartment(s) is affected by the transfection method (transient expressionversusstable expression) and suggested that Syt VII molecule on dense-core vesicles functions as a vesicular Ca2+sensor for exocytosis in endocrine cells.

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