Proteomics

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SILAC-based Quantitative Proteomic Analysis Unveiled the Arsenite-induced Perturbation of Multiple Pathways in Human Skin Fibroblast Cells


ABSTRACT: Humans are exposed to arsenic species through inhalation, ingestion and dermal contact, which may lead to skin, liver, and bladder cancers as well as cardiovascular and neurological diseases. The mechanisms underlying the cytotoxic and carcinogenic effects of arsenic species, however, remain incompletely understood. To exploit the mechanisms of toxicity of As(III), we employed stable isotope labeling by amino acids in cell culture (SILAC) together with LC-MS/MS analysis to assess quantitatively the As(III)-induced perturbation of the entire proteome of cultured human skin fibroblast cells. Shotgun proteomic analysis on an LTQ-Orbitrap Velos mass spectrometer facilitated the quantification of more than 3800 proteins, 250 of which displayed significant alternations (> 1.5 fold) upon arsenite treatment. Targeted analysis on a triple-quadrupole mass spectrometer in the multiple-reaction monitoring (MRM) mode confirmed the quantification results of some select proteins. Ingenuity pathway analysis revealed the arsenite-induced alteration of more than 10 biological pathways, including the Nrf2-mediated oxidative stress response pathway as reflected by the up-regulation of 9 proteins in this pathway. In addition, arsenite induced changes in expression levels of a number of selenoproteins and metallothioneins. Together, the results from the present study painted a more complete picture about the biological pathways that are altered in human skin fibroblast cells upon arsenite exposure.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Fibroblast

SUBMITTER: Fan Zhang  

LAB HEAD: Fan Zhang

PROVIDER: PXD005646 | Pride | 2018-10-26

REPOSITORIES: Pride

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Publications

SILAC-Based Quantitative Proteomic Analysis Unveils Arsenite-Induced Perturbation of Multiple Pathways in Human Skin Fibroblast Cells.

Zhang Fan F   Xiao Yongsheng Y   Wang Yinsheng Y  

Chemical research in toxicology 20170214 4


Humans are exposed to arsenic species through inhalation, ingestion, and dermal contact, which may lead to skin, liver, and bladder cancers as well as cardiovascular and neurological diseases. The mechanisms underlying the cytotoxic and carcinogenic effects of arsenic species, however, remain incompletely understood. To exploit the mechanisms of toxicity of As(III), we employed stable isotope labeling by amino acids in cell culture (SILAC) together with LC/MS/MS analysis to quantitatively assess  ...[more]

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