Proteomics Analysis Reveals Involvement of Krt17 in Areca Nut

Jul 19, 2016 - Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Kwei-Shan, Tao-Yuan 333, Taiwan...
2 downloads 6 Views 3MB Size
Subscriber access provided by RMIT University Library

Article

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

Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a free service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are accessible to all readers and citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.

Journal of Proteome Research is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.

Page 1 of 54

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

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

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

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