Pediatric Surgery International

, Volume 29, Issue 11, pp 1147–1152 | Cite as

RASSF1A methylation indicates a poor prognosis in hepatoblastoma patients

  • Shohei Honda
  • Hisayuki Miyagi
  • Hiromu Suzuki
  • Masashi Minato
  • Masayuki Haruta
  • Yasuhiko Kaneko
  • Kanako C. Hatanaka
  • Eiso Hiyama
  • Takehiko Kamijo
  • Tadao OkadaEmail author
  • Akinobu Taketomi
Original Article



The RAS association domain family protein 1 (RASSF1A) is known to be frequently inactivated by promoter hypermethylation in cancers. This study investigated the association of RASSF1A methylation with clinical outcomes in hepatoblastoma patients and whether it is correlated with the histological phenotype of hepatoblastoma tumors.


Seventy-four hepatoblastoma tumors were obtained from patients enrolled in the Japanese study group for pediatric liver tumor protocol-2. From nine formalin-fixed, paraffin-embedded specimens, we extracted DNA by dissection under a light microscope. We examined the methylation status of the RASSF1A promoter region by bisulfite pyrosequencing.


Twenty-five (33.8 %) hepatoblastoma tumors were classified as having methylated RASSF1A. The RASSF1A methylation was significantly associated with metastatic tumors and a poor prognosis. Despite the complete resection, five pretreatment extent of disease II tumors showed recurrence or distant metastasis postoperatively. Among these cases, four tumors were found to show RASSF1A methylation. When compared to histologically different types of cell, RASSF1A methylation values in samples of the normal liver, fetal type, and embryonal type, were significantly elevated in ascending order.


We confirmed that RASSF1A methylation is a significant prognostic indicator in hepatoblastomas, and it may become a promising molecular marker to stratify patients into appropriate risk groups.


Hepatoblastoma RASSF1A methylation Prognostic marker 



We would like to thank all the staff at institutes that participated in JPLT for enrolling their patients in the study. We are also grateful to JPLT steering committee members (Drs. T. Hishiki, K. Ida, K. Watanabe, S. Kondo, T. Oue, M. Yano, and T. Tajiri), JPLT pathological committee members (Drs. H. Horie, Y. Tanaka, and K. Inoue) and a data administrator for JPLT (Dr. K. Hiyama, Hiroshima University), for data managements and clinic-pathological review of these patients.

Conflict of interest

The authors who have taken part in this study declare that they do not have anything to disclose regarding funding or any conflict of interest with respect to this manuscript. The first and the corresponding authors are JSPS members, and this abstract was selected for presentation at the 50th Annual Meeting of the Japanese Society of Pediatric Surgeons.


  1. 1.
    von Schweinitz D (2012) Hepatoblastoma: recent developments in research and treatment. Semin Pediatr Surg 21:21–30CrossRefGoogle Scholar
  2. 2.
    Zimmermann A, Lopez-Terrada D (2011) Pathology of pediatric liver tumors. In: Zimmermann A, Perilongo G, Malogolowkin M, von Schweinitz D (eds) Pediatric liver tumors. Springer, Heidelberg, pp 83–112CrossRefGoogle Scholar
  3. 3.
    Agathanggelou A, Cooper NC, Latif F et al (2005) Role of the Ras-association domain family 1 tumor suppressor gene in human cancers. Cancer Res 65:3497–3508PubMedCrossRefGoogle Scholar
  4. 4.
    Honda S, Haruta M, Sugawara W et al (2008) The methylation status of RASSF1A promoter predicts responsiveness to chemotherapy and eventual cure in hepatoblastoma patients. Int J Cancer 123:1117–1125PubMedCrossRefGoogle Scholar
  5. 5.
    Hishiki T, Matsunaga T, Sasaki F et al (2011) Outcome of hepatoblastomas treated using the Japanese study group for pediatric liver tumor (JPLT) protocol-2: report from the JPLT. Pediatr Surg Int 27:1–8PubMedCrossRefGoogle Scholar
  6. 6.
    Brown J, Perilongo G, Shafford E et al (2000) Pretreatment prognostic factors for children with hepatoblastoma- results from the International society of paediatric oncology (SIOP) study SIOPEL 1. Eur J Cancer 36:1418–1425PubMedCrossRefGoogle Scholar
  7. 7.
    Yang AS, Estécio MR, Doshi K et al (2004) A simple method for estimating global DNA methylation using bisulfite PCR of repetitive DNA elements. Nucleic Acids Res 32:e38PubMedCrossRefGoogle Scholar
  8. 8.
    Ueda Y, Hiyama E, Kamimatsuse A et al (2011) Wnt signaling and telomerase activation of hepatoblastoma: correlation with chemosensitivity and surgical resectability. J Pediatr Surg 46:2221–2227PubMedCrossRefGoogle Scholar
  9. 9.
    Yamada S, Ohira M, Horie H et al (2004) Expression profiling and differential screening between hepatoblastomas and the corresponding normal livers: identification of high expression of the PLK1 oncogene as a poor-prognostic indicator of hepatoblastomas. Oncogene 23:5901–5911PubMedCrossRefGoogle Scholar
  10. 10.
    Agathanggelou A, Cooper WN, Latif F (2005) Role of the Ras-association domain family 1 tumor suppressor gene in human cancers. Cancer Res 65:3497–3508PubMedCrossRefGoogle Scholar
  11. 11.
    Yang Q, Zage P, Kagan D et al (2004) Association of epigenetic inactivation of RASSF1A with poor outcome in human neuroblastoma. Clin Cancer Res 10:8493–8500PubMedCrossRefGoogle Scholar
  12. 12.
    Ohshima J, Haruta M, Fujiwara Y et al (2012) Methylation of the RASSF1A promoter is predictive of poor outcome among patients with Wilms tumor. Pediatr Blood Cancer 59:499–505PubMedCrossRefGoogle Scholar
  13. 13.
    Cairo S, Armengol C, De Reyniès A et al (2008) Hepatic stem-like phenotype and interplay of Wnt/beta-catenin and Myc signaling in aggressive childhood liver cancer. Cancer Cell 14:471–484PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Shohei Honda
    • 1
  • Hisayuki Miyagi
    • 1
  • Hiromu Suzuki
    • 2
  • Masashi Minato
    • 1
  • Masayuki Haruta
    • 3
  • Yasuhiko Kaneko
    • 3
  • Kanako C. Hatanaka
    • 4
  • Eiso Hiyama
    • 5
  • Takehiko Kamijo
    • 6
  • Tadao Okada
    • 1
    Email author
  • Akinobu Taketomi
    • 1
  1. 1.Department of Gastroenterological Surgery IHokkaido University Graduate School of MedicineSapporoJapan
  2. 2.Department of Molecular BiologySapporo Medical UniversitySapporoJapan
  3. 3.Department of Cancer Diagnosis, Research Institute for Clinical OncologySaitama Cancer CenterSaitamaJapan
  4. 4.Department of Surgical PathologyHokkaido University HospitalSapporoJapan
  5. 5.Japanese Study Group for Pediatric Liver TumorHiroshimaJapan
  6. 6.Department of Biochemistry and Molecular CarcinogenesisChiba Cancer Center Research InstituteChibaJapan

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