title: SILAC-Based Quantitative Proteomic Analysis Unveils 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 quantitatively assess 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 3880 proteins, 130 of which were quantified in both forward and reverse SILAC-labeling experiments and displayed significant alterations (>1.5 fold) upon arsenite treatment. Targeted analysis on a triple-quadrupole mass spectrometer in 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, which is represented by the upregulation of nine 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 regarding the biological pathways that are altered in human skin fibroblast cells upon arsenite exposure.
authors.affiliation: Department of Chemistry, University of California , Riverside, California 92521-0403, United States.
authors.forename: Fan
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Department of Chemistry, University of California , Riverside, California 92521-0403, United States.
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: Department of Chemistry, University of California , Riverside, California 92521-0403, United States.
authors.forename: Yinsheng
authors.identifier: 0000-0001-5565-283X
authors.initials: Y
authors.lastname: Wang
mesh_terms: D018053:Arsenites / Q000633:toxicity*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D015536:Down-Regulation / Q000187:drug effects; D005293:Ferritins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D007553:Isotope Labeling; D008668:Metallothionein / Q000378:metabolism; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D020543:Proteome / Q000032:analysis / Q000737:chemistry / Q000187:drug effects*; D040901:Proteomics; D017382:Reactive Oxygen Species / Q000378:metabolism; D051140:Selenoproteins / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D053719:Tandem Mass Spectrometry; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D018053:Arsenites; D016209:Interleukin-8; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D020543:Proteome; D017382:Reactive Oxygen Species; D051140:Selenoproteins; D005293:Ferritins; D008668:Metallothionein; C015001:arsenite