title: Cd²⁺-induced alteration of the global proteome of human skin fibroblast cells.
abstract: Cadmium (Cd(2+)) is a toxic heavy metal and a well-known human carcinogen. The toxic effects of Cd(2+) on biological systems are diverse and thought to be exerted through a complex array of mechanisms. Despite the large number of studies aimed to elucidate the toxic mechanisms of action of Cd(2+), few have been targeted toward investigating the ability of Cd(2+) to disrupt multiple cellular pathways simultaneously and the overall cellular responses toward Cd(2+) exposure. In this study, we employed a quantitative proteomic method, relying on stable isotope labeling by amino acids in cell culture (SILAC) and LC-MS/MS, to assess the Cd(2+)-induced simultaneous alterations of multiple cellular pathways in cultured human skin fibroblast cells. By using this approach, we were able to quantify 2931 proteins, and 400 of them displayed significantly changed expression following Cd(2+) exposure. Our results unveiled that Cd(2+) treatment led to the marked upregulation of several antioxidant enzymes (e.g., metallothionein-1G, superoxide dismutase, pyridoxal kinase, etc.), enzymes associated with glutathione biosynthesis and homeostasis (e.g., glutathione S-transferases, glutathione synthetase, glutathione peroxidase, etc.), and proteins involved in cellular energy metabolism (e.g., glycolysis, pentose phosphate pathway, and the citric acid cycle). Additionally, we found that Cd(2+) treatment resulted in the elevated expression of two isoforms of dimethylarginine dimethylaminohydrolase (DDAH I and II), enzymes known to play a key role in regulating nitric oxide biosynthesis. Consistent with these findings, we observed elevated formation of nitric oxide in human skin (GM00637) and lung (IMR-90) fibroblast cells following Cd(2+) exposure. The upregulation of DDAH I and II suggests a role of nitric oxide synthesis in Cd(2+)-induced toxicity in human cells.
authors: Department of Chemistry and ‡Environmental Toxicology Graduate Program, University of California , Riverside, California 92521-0403, United States.
authors: John M
authors: JM
authors: Prins
authors: Lijuan
authors: L
authors: Fu
authors: Lei
authors: L
authors: Guo
authors: Yinsheng
authors: Y
authors: Wang
mesh_terms: D000581:Amidohydrolases / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D019256:Cadmium Chloride / Q000633:toxicity*; D002460:Cell Line; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D007553:Isotope Labeling; D008168:Lung / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D053858:Metabolic Networks and Pathways / Q000187:drug effects; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000819:agonists* / Q000096:biosynthesis; D025941:Protein Interaction Mapping; D020543:Proteome / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D012867:Skin / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
chemical_list: D020543:Proteome; D009569:Nitric Oxide; D000581:Amidohydrolases; C059999:dimethylargininase; D019256:Cadmium Chloride