Kiminobu Sugito and Hiroyuki Kawashima contributed equally to this study.
Article first published online: 21 AUG 2012
Copyright © 2012 Wiley Periodicals, Inc.
Pediatric Blood & Cancer
Volume 60, Issue 3, pages 383–389, March 2013
How to Cite
Sugito, K., Kawashima, H., Uekusa, S., Yoshizawa, S., Hoshi, R., Furuya, T., Kaneda, H., Hosoda, T., Masuko, T., Ohashi, K., Ikeda, T., Koshinaga, T., Fujiwara, K., Igarashi, J., Ghosh, S., Held, W. A. and Nagase, H. (2013), Identification of aberrant methylation regions in neuroblastoma by screening of tissue-specific differentially methylated regions. Pediatr. Blood Cancer, 60: 383–389. doi: 10.1002/pbc.24282
Authors' contributions: KS, HK, SU, SY, and RH carried out the methylation analysis using the MassARRAY EpiTYPER system. KS, TF, HK, TH, TM, TI, and TK collected the clinical samples and data. KF, JI, SG, WA, and HN were involved in the conceptual design of the study. HN was responsible for the supervision of the project. KS, HK, and HN wrote the manuscript. All authors read and approved the final manuscript.
Conflict of interest: Nothing to declare.
- Issue published online: 15 JAN 2013
- Article first published online: 21 AUG 2012
- Manuscript Accepted: 12 JUL 2012
- Manuscript Received: 13 MAR 2012
- Nihon University Multidisciplinary Research Grant for 2006, 2007
- Academic Frontier Project for 2006 Project for Private Universities
- National Cancer Institute Grant. Grant Number: CA102423
- National Cancer Institute Center Support Grant. Grant Number: CA16056
- Ministry of Education, Science, Sports, and Culture of Japan. Grant Number: 20592092
- prognostic factors;
- tissue-specific differentially methylated regions
The identification of tissue-specific differentially methylated regions (tDMRs) is key to our understanding of mammalian development. Research has indicated that tDMRs are aberrantly methylated in cancer and may affect the oncogenic process.
We used the MassARRAY EpiTYPER system to determine the quantitative methylation levels of seven neuroblastomas (NBs) and two control adrenal medullas at 12 conserved tDMRs. A second sample set of 19 NBs was also analyzed. Statistical analysis was carried out to determine the relationship of the quantitative methylation levels to other prognostic factors in these sample sets.
Screening of 12 tDMRs revealed 2 genomic regions (SLC16A5 and ZNF206) with frequent aberrant methylation patterns in NB. The methylation levels of SLC16A5 and ZNF206 were low compared to the control adrenal medullas. The SLC16A5 methylation level (cut-off point, 13.25%) was associated with age at diagnosis, disease stage, and Shimada classification but not with MYCN amplification. The ZNF206 methylation level (cut-off point, 68.80%) was associated with all of the prognostic factors analyzed. Although the methylation levels at these regions did not reach statistical significance in their association with prognosis in mono-variant analysis, patients with both hypomethylation of SLC16A5 and hypermethylation of ZNF206 had a significantly prolonged event-free survival, when these two variables were analyzed together.
We demonstrated that two tDMRs frequently displayed altered methylation patterns in the NB genome, suggesting their distinct involvement in NB development/differentiation. The combined analysis of these two regions could serve as a diagnostic biomarker for poor clinical outcome. Pediatr Blood Cancer 2013; 60: 383–389. © 2012 Wiley Periodicals, Inc.