title: SILAC-based quantitative proteomic analysis reveals widespread molecular alterations in human skin keratinocytes upon chronic arsenic exposure.
abstract: Chronic exposure to arsenic is associated with dermatological and nondermatological disorders. Consumption of arsenic-contaminated drinking water results in accumulation of arsenic in liver, spleen, kidneys, lungs, and gastrointestinal tract. Although arsenic is cleared from these sites, a substantial amount of residual arsenic is left in keratin-rich tissues including skin. Epidemiological studies suggest the association of skin cancer upon arsenic exposure, however, the mechanism of arsenic-induced carcinogenesis is not completely understood. We developed a cell line based model to understand the molecular mechanisms involved in arsenic-mediated toxicity and carcinogenicity. Human skin keratinocyte cell line, HaCaT, was chronically exposed to 100 nM sodium arsenite over a period of 6 months. We observed an increase in basal ROS levels in arsenic-exposed cells. SILAC-based quantitative proteomics approach resulted in identification of 2111 proteins of which 42 proteins were found to be overexpressed and 54 downregulated (twofold) upon chronic arsenic exposure. Our analysis revealed arsenic-induced overexpression of aldo-keto reductase family 1 member C2 (AKR1C2), aldo-keto reductase family 1 member C3 (AKR1C3), glutamate-cysteine ligase catalytic subunit (GCLC), and NAD(P)H dehydrogenase [quinone] 1 (NQO1) among others. We observed downregulation of several members of the plakin family including periplakin (PPL), envoplakin (EVPL), and involucrin (IVL) that are essential for terminal differentiation of keratinocytes. MRM and Western blot analysis confirmed differential expression of several candidate proteins. Our study provides insights into molecular alterations upon chronic arsenic exposure on skin.
authors.affiliation: Institute of Bioinformatics, International Technology Park, Bangalore, India.|Manipal University, Manipal, Karnataka, India.
authors.forename: Sartaj Ahmad
authors.initials: SA
authors.lastname: Mir
authors.affiliation: Institute of Bioinformatics, International Technology Park, Bangalore, India.|YU-IOB Center for Systems Biology and Molecular Medicine, Yenepoya University, Mangalore, India.
authors.forename: Sneha M
authors.initials: SM
authors.lastname: Pinto
authors.affiliation: Molecular Genetics Division, CSIR-Indian Institute of Chemical Biology, Kolkata, India.
authors.forename: Somnath
authors.initials: S
authors.lastname: Paul
authors.affiliation: Institute of Bioinformatics, International Technology Park, Bangalore, India.
authors.forename: Remya
authors.initials: R
authors.lastname: Raja
authors.affiliation: Institute of Bioinformatics, International Technology Park, Bangalore, India.|Amrita School of Biotechnology, Amrita University, Kollam, India.
authors.forename: Vishalakshi
authors.initials: V
authors.lastname: Nanjappa
authors.affiliation: Institute of Bioinformatics, International Technology Park, Bangalore, India.|Department of Biochemistry and Molecular Biology, Pondicherry University, Puducherry, India.
authors.forename: Nazia
authors.initials: N
authors.lastname: Syed
authors.affiliation: Institute of Bioinformatics, International Technology Park, Bangalore, India.|Manipal University, Manipal, Karnataka, India.
authors.forename: Jayshree
authors.initials: J
authors.lastname: Advani
authors.affiliation: Institute of Bioinformatics, International Technology Park, Bangalore, India.
authors.forename: Santosh
authors.initials: S
authors.lastname: Renuse
authors.affiliation: Institute of Bioinformatics, International Technology Park, Bangalore, India.
authors.forename: Nandini A
authors.initials: NA
authors.lastname: Sahasrabuddhe
authors.affiliation: Institute of Bioinformatics, International Technology Park, Bangalore, India.|YU-IOB Center for Systems Biology and Molecular Medicine, Yenepoya University, Mangalore, India.|NIMHANS-IOB Proteomics and Bioinformatics Laboratory, Neurobiology Research Centre, National Institute of Mental Health and Neurosciences, Bangalore, India.
authors.forename: T S Keshava
authors.initials: TS
authors.lastname: Prasad
authors.affiliation: Molecular Genetics Division, CSIR-Indian Institute of Chemical Biology, Kolkata, India.
authors.forename: Ashok K
authors.initials: AK
authors.lastname: Giri
authors.affiliation: Institute of Bioinformatics, International Technology Park, Bangalore, India.|YU-IOB Center for Systems Biology and Molecular Medicine, Yenepoya University, Mangalore, India.
authors.forename: Harsha
authors.initials: H
authors.lastname: Gowda
authors.affiliation: Institute of Bioinformatics, International Technology Park, Bangalore, India.|YU-IOB Center for Systems Biology and Molecular Medicine, Yenepoya University, Mangalore, India.
authors.forename: Aditi
authors.initials: A
authors.lastname: Chatterjee
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism*; D001151:Arsenic / Q000633:toxicity*; D015153:Blotting, Western; D002460:Cell Line; D019295:Computational Biology; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D000072019:Kelch-Like ECH-Associated Protein 1 / Q000378:metabolism; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism*; D051267:NF-E2-Related Factor 2 / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D020543:Proteome / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D015203:Reproducibility of Results; D015398:Signal Transduction / Q000187:drug effects; D012867:Skin / Q000166:cytology*
chemical_list: D000596:Amino Acids; C431364:KEAP1 protein, human; D000072019:Kelch-Like ECH-Associated Protein 1; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D020543:Proteome; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; D001151:Arsenic
keywords: Arsenic poisoning; Biomedicine; Keratinocytes; Metabolic labeling