title: Proteomics Analysis Reveals Involvement of Krt17 in Areca Nut-Induced Oral Carcinogenesis.
abstract: The areca nut is a known carcinogen that causes oral cancer in individuals in Southeast Asia, but the molecular mechanism that leads to this malignancy is still unclear. To mimic the habit of areca nut chewing, our laboratory has established four oral cancer cell sublines (SAS, OECM1, K2, C9), which have been chronically exposed to areca nut extract (ANE). To elucidate the molecular basis of areca nut-induced oral carcinogenesis, the differential proteomes between oral cancer cells and the ANE-treated sublines were determined using isobaric mass tag (iTRAQ) labeling and multidimensional liquid chromatography-mass spectrometry (LC-MS/MS). Over 1000 proteins were identified in four sublines, and 196 proteins were found to be differentially expressed in at least two ANE-treated sublines. A bioinformatic analysis revealed that these proteins participate in several pathways, and one of the most prominent pathways was the regulation of epithelial to mesenchymal transition (EMT). In all, 24 proteins including Krt17 were confirmed to be differentially expressed in the ANE-treated sublines. To reveal additional information on the mechanism of ANE-induced carcinogenesis, Krt17 was further investigated. Krt17 knockdown significantly suppressed ANE-induced cell migration and invasion and modulated the EMT process. Furthermore, in a murine model of carcinogen-induced (arecoline cocktail, an active compound of ANE) oral cancer, Krt17 was significantly up-regulated in all hyperplastic tissues and in carcinoma tissues (p < 0.001). In conclusion, we have identified a proteome of oral cancer cells that is associated with chronic areca nut exposure. Krt17 was demonstrated to contribute to areca nut-induced oral malignancy. The results of this study contribute to risk assessment, disease prevention and other clinical applications associated with areca nut-induced oral cancer.
authors.affiliation: Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University , Kwei-Shan, Tao-Yuan 333, Taiwan.|Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University , Kwei-Shan, Tao-Yuan 333, Taiwan.
authors.forename: Chang-Hsu
authors.initials: CH
authors.lastname: Chiang
authors.affiliation: Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University , Kwei-Shan, Tao-Yuan 333, Taiwan.|Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University , Kwei-Shan, Tao-Yuan 333, Taiwan.|Molecular Medicine Research Center, Chang Gung University , Kwei-Shan, Tao-Yuan 333, Taiwan.|Department of Otolaryngology-Head & Neck Surgery, Chang Gung Memorial Hospital , Linkou 333, Taiwan.
authors.forename: Chih-Ching
authors.initials: CC
authors.lastname: Wu
authors.affiliation: Department of Pathology, Chang Gung Memorial Hospital , Tao-Yuan 333, Taiwan.
authors.forename: Li-Yu
authors.initials: LY
authors.lastname: Lee
authors.affiliation: Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University , Kwei-Shan, Tao-Yuan 333, Taiwan.
authors.forename: Yi-Chen
authors.initials: YC
authors.lastname: Li
authors.affiliation: Department of Veterinary Medicine, National Chung Hsing University , Tai-Chung 402, Taiwan.
authors.forename: Hao-Ping
authors.initials: HP
authors.lastname: Liu
authors.affiliation: Molecular Medicine Research Center, Chang Gung University , Kwei-Shan, Tao-Yuan 333, Taiwan.
authors.forename: Chia-Wei
authors.initials: CW
authors.lastname: Hsu
authors.affiliation: Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University , Kwei-Shan, Tao-Yuan 333, Taiwan.
authors.forename: Ya-Ching
authors.initials: YC
authors.lastname: Lu
authors.affiliation: Department of Radiation Oncology, Chang Gung Memorial Hospital , Tao-Yuan 333, Taiwan.
authors.forename: Joseph T
authors.initials: JT
authors.lastname: Chang
authors.affiliation: Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University , Kwei-Shan, Tao-Yuan 333, Taiwan.|Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University , Kwei-Shan, Tao-Yuan 333, Taiwan.
authors.forename: Ann-Joy
authors.initials: AJ
authors.lastname: Cheng
mesh_terms: D000818:Animals; D001114:Areca / Q000737:chemistry / Q000633:toxicity*; D002460:Cell Line; D019295:Computational Biology; D058750:Epithelial-Mesenchymal Transition; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D053537:Keratin-17 / Q000378:metabolism* / Q000502:physiology; D051379:Mice; D009062:Mouth Neoplasms / Q000209:etiology*; D010936:Plant Extracts / Q000494:pharmacology*; D040901:Proteomics / Q000379:methods*; D014407:Tumor Cells, Cultured
chemical_list: D053537:Keratin-17; D010936:Plant Extracts
keywords: Krt17; areca nut; epithelial to mesenchymal transition; iTRAQ; oral carcinogenesis