Retrospective Study
Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Apr 7, 2015; 21(13): 3960-3969
Published online Apr 7, 2015. doi: 10.3748/wjg.v21.i13.3960
Methylation of IRAK3 is a novel prognostic marker in hepatocellular carcinoma
Chih-Chi Kuo, Yu-Lueng Shih, Her-Young Su, Ming-De Yan, Chung-Bao Hsieh, Chin-Yu Liu, Wei-Ting Huang, Mu-Hsien Yu, Ya-Wen Lin
Chih-Chi Kuo, Ya-Wen Lin, Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei 114, Taiwan
Yu-Lueng Shih, Division of Gastroenterology, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei 114, Taiwan
Her-Young Su, Mu-Hsien Yu, Department of Obstetrics and Gynecology, Tri-Service General Hospital, National Defense Medical Center, Taipei 114, Taiwan
Ming-De Yan, Cancer Center, Wan Fang Hospital, Taipei Medical University, Taipei 116, Taiwan
Chung-Bao Hsieh, Division of General Surgery, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei 114, Taiwan
Chin-Yu Liu, Department of Nutritional Science, Fu Jen Catholic University, New Taipei City 242, Taiwan
Wei-Ting Huang, Ya-Wen Lin, Department and Graduate Institute of Microbiology and Immunology, Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei 114, Taiwan
Ya-Wen Lin, Graduate Institute of Life Sciences, National Defense Medical Center, Taipei 114, Taiwan
Author contributions: Kuo CC, Shih YL and Huang WT performed the majority of experiments; Shih YL, Su HY, Yan MD, Hsieh CB, Liu CY and Yu MH provided vital reagents and analytical tools and also helped to edit the manuscript; Shih YL, Hsieh CB and Yu MH coordinated and provided the collection of all the human material and also provided financial support for this work; Kuo CC and Lin YW designed the study and wrote the manuscript.
Supported by National Science Council, No. NSC 102-2320-B-016-016-MY3, Taiwan; and the Liver Disease Prevention and Treatment Research Foundation, Taiwan.
Ethics approval: The study was reviewed and approved by the Institutional Review Board of the Tri-Service General Hospital and TLCN User Committee.
Informed consent: Not applicable. This is a delinked tissue bank of Taiwan. Researchers can apply samples for study after the approval of TLCN User Committee and Institutional Review Board of the Tri-Service General Hospital.
Conflict-of-interest: A conflict-of-interest statement is included in the manuscript.
Data sharing: No additional data are available.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Ya-Wen Lin, PhD, Department and Graduate Institute of Microbiology and Immunology, Graduate Institute of Medical Sciences, National Defense Medical Center, No. 161, Section 6, Min-Chuan East Road, Taipei 114, Taiwan. ndmc.yawen@msa.hinet.net
Telephone: +886-2-87917654 Fax: +886-2-87917654
Received: September 3, 2014
Peer-review started: September 4, 2014
First decision: September 27, 2014
Revised: November 7, 2014
Accepted: December 14, 2014
Article in press: December 16, 2014
Published online: April 7, 2015
Processing time: 216 Days and 3.7 Hours
Abstract

AIM: To examine the methylation levels of interleukin-1 receptor-associated kinase 3 (IRAK3) and GLOXD1 and their potential clinical applications in hepatocellular carcinoma (HCC).

METHODS: mRNA expression and promoter methylation of IRAK3 and GLOXD1 in HCC cells were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and methylation-specific PCR (MSP), respectively. Using pyrosequencing results, we further established a quantitative MSP (Q-MSP) system for the evaluation of IRAK3 and GLOXD1 methylation in 29 normal controls and 160 paired HCC tissues and their adjacent nontumor tissues. We also calculated Kaplan-Meier survival curves to determine the applications of gene methylation in the prognosis of HCC.

RESULTS: IRAK3 and GLOXD1 expression was partially restored in several HCC cell lines after treatment with 5-aza-2′-deoxycytidine (DNA methyltransferase inhibitor; 5DAC). A partial decrease in the methylated band was also observed in the HCC cell lines after 5DAC treatment. Using GLOXD1 as an example, we found a significant correlation between the data obtained from the methylation array and from pyrosequencing. The methylation frequency of IRAK3 and GLOXD1 in HCC tissues was 46.9% and 63.8%, respectively. Methylation of IRAK3 was statistically associated with tumor stage. Moreover, HCC patients with IRAK3 methylation had a trend toward poor 3-year disease-free survival (P < 0.05).

CONCLUSION: IRAK3 and GLOXD1 were frequently methylated in HCC tissues compared to normal controls and nontumor tissues. IRAK3 methylation was associated with tumor stage and poor prognosis of patients. These data suggest that IRAK3 methylation is a novel prognostic marker in HCC.

Keywords: IRAK3; GLOXD1; Hepatocellular carcinoma; DNA methylation biomarker; Quantitative methylation-specific polymerase chain reaction; Pyrosequencing

Core tip: The methylation biomarker is relatively stable in tissue samples and body fluids, suggesting that it is a good tool for the detection, diagnosis, prognosis, and even therapy of hepatocellular carcinoma (HCC). Our study not only demonstrated frequent methylation of interleukin-1 receptor-associated kinase 3 (IRAK3) and GLOXD1 in HCC but also found that IRAK3 methylation was positively associated with poor 3-year disease-free survival of patients. This indicates that IRAK3 methylation could be used as a potential biomarker for prediction of prognosis in HCC.