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Proteomics analysis reveals involvement of Krt17 in areca nut-induced oral carcinogenesis Chang-Hsu Chiang, Chih-Ching Wu, Li-Yu Lee, Yi-Chen Li, Hao-Ping Liu, Chia-Wei Hsu, Ya-Ching Lu, Joseph T. Chang, and Ann-Joy Cheng J. Proteome Res., Just Accepted Manuscript • DOI: 10.1021/acs.jproteome.6b00138 • Publication Date (Web): 19 Jul 2016 Downloaded from http://pubs.acs.org on July 21, 2016
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Journal of Proteome Research
Proteomics analysis reveals involvement of Krt17 in areca nut-induced oral carcinogenesis
Chang-Hsu Chianga,b,#, Chih-Ching Wua,b,c,d,#, Li-Yu Leee, Yi-Chen Lia, Hao-Ping Liuf, Chia-Wei Hsuc, Ya-Ching Lua, Joseph T. Changg,*, Ann-Joy Chenga,b,*
a
Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung
University, Kwei-Shan, Tao-Yuan, Taiwan b
Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University,
Kwei-Shan, Tao-Yuan, Taiwan c
Molecular Medicine Research Center, Chang Gung University, Kwei-Shan, Tao-Yuan, Taiwan
d
Department of Otolaryngology-Head & Neck Surgery, Chang Gung Memorial Hospital, Linkou,
Taiwan e
Department of Pathology, Chang Gung Memorial Hospital, Tao-Yuan, Taiwan
f
Department of Veterinary Medicine, National Chung Hsing University, Tai-Chung, Taiwan
g
Department of Radiation Oncology, Chang Gung Memorial Hospital, Tao-Yuan, Taiwan
#
First and second authors contributed equally.
* Correspondence to Dr. Ann-Joy Cheng, Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, 259 Wen-Hwa 1st Road, Tao-Yuan 333, Taiwan. Tel: 886-3-2118800 ex 5085, Fax: 886-3-2118247, e-mail:
[email protected]; or Dr. Joseph T. Chang, Department of Radiation Oncology, Chang Gung Memorial Hospital, 5 Fu-Shin Road, Tao-Yuan 333, Taiwan. Tel: 886-3-3281200 ex 7008, Fax; 886-3-2118247, e-mail:
[email protected].
Running title: iTRAQ-based discovery of Krt17 involvement in areca nut-induced oral carcinogenesis
ACS Paragon Plus 1 Environment
Journal of Proteome Research
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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 one thousand 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 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
