Annals of Surgical Oncology

, Volume 18, Issue 2, pp 572–579 | Cite as

Reduced Expression of Reelin (RELN) Gene Is Associated With High Recurrence Rate of Hepatocellular Carcinoma

  • Yukiyasu Okamura
  • Shuji Nomoto
  • Mitsuro Kanda
  • Masamichi Hayashi
  • Yoko Nishikawa
  • Tsutomu Fujii
  • Hiroyuki Sugimoto
  • Shin Takeda
  • Akimasa Nakao
Translational Research and Biomarkers



Hepatocellular carcinoma (HCC) is one of the world’s top five causes of cancer-related deaths. Current treatments available ameliorate HCC; however, current therapy fails to completely treat and prevent HCC, as shown by its high recurrence rate. Recently developed genome-wide gene-expression profile analyses can now robustly detect many candidate genes that are modified by HCC. Here we attempt to identify novel genes displaying altered gene expression profiles when comparing healthy tissue with HCC by means of a double-combination array previously developed.


Double-combination array analysis of gene expression profiles and single nucleotide polymorphism arrays were performed on each HCC tissue sample. Subsequently, samples from 48 HCC patients were subjected to quantitative real-time reverse transcription polymerase chain reaction and methylation-specific polymerase chain reaction.


The reelin (RELN) gene was detected as a pertinent tumor suppressor gene by means of this method. Of the 48 clinical samples obtained, 34 (79.2%) displayed reduced RELN expression in tumor tissue, and the expression level of tumor tissues clearly reduced compared with that of corresponding normal tissues (P = 0.002). Eighteen (37.5%) of 48 tumor tissues were found to be hypermethylated on the RELN gene promoter. Moreover, analysis of clinical data revealed an inverse correlation between RELN expression and HCC recurrence.


The present study indicates that our in-house double-combination array is an effective and convenient technique in detecting novel genes with altered expression in disease. We suggest RELN is a key regulatory gene associated with the recurrence of HCC.


Single Nucleotide Polymorphism Promoter Hypermethylation Leukemia Inhibitory Factor Receptor RELN Gene Single Nucleotide Polymorphism Array 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Conflict of interest

The authors declare no conflict of interest.


  1. 1.
    Parkin DM, Bray F, Ferlay J, Pisani P. Global cancer statistics, 2002. CA Cancer J Clin. 2005;55:74–108.CrossRefPubMedGoogle Scholar
  2. 2.
    Arii S, Okamoto E, Imamura M. Registries in Japan: current status of hepatocellular carcinoma in Japan. Liver Cancer Study Group of Japan. Semin Surg Oncol. 1996;12:204–11.Google Scholar
  3. 3.
    Thorgeirsson SS, Grisham JW. Molecular pathogenesis of human hepatocellular carcinoma. Nat Genet. 2002;31:339–46.CrossRefPubMedGoogle Scholar
  4. 4.
    Hsu IC, Metcalf RA, Sun T, et al. Mutational hotspot in the p53 gene in human hepatocellular carcinomas. Nature. 1991;350:427–8.CrossRefPubMedGoogle Scholar
  5. 5.
    Zhang X, Xu HJ, Murakami Y, et al. Deletions of chromosome 13q, mutations in retinoblastoma 1, and retinoblastoma protein state in human hepatocellular carcinoma. Cancer Res. 1994;54:4177–82.PubMedGoogle Scholar
  6. 6.
    Yamada T, de Souza AT, Finkelstein S, Jirtle RL. Loss of the gene encoding mannose 6-phosphate/insulin-like growth factor II receptor is an early event in liver carcinogenesis. Proc Natl Acad Sci USA. 1997;94:10351–5.CrossRefPubMedGoogle Scholar
  7. 7.
    Zhang XK, Huang DP, Qiu DK, Chiu JF. The expression of c-myc and c-N-ras in human cirrhotic livers, hepatocellular carcinomas and liver tissue surrounding the tumors. Oncogene. 1990;5:909–14.PubMedGoogle Scholar
  8. 8.
    Nishida N, Fukuda Y, Komeda T, et al. Amplification and overexpression of the cyclin D1 gene in aggressive human hepatocellular carcinoma. Cancer Res. 1994;54:3107–10.PubMedGoogle Scholar
  9. 9.
    Hsu HC, Jeng YM, Mao TL, et al. Beta-catenin mutations are associated with a subset of low-stage hepatocellular carcinoma negative for hepatitis B virus and with favorable prognosis. Am J Pathol. 2000;157:763–70.CrossRefPubMedGoogle Scholar
  10. 10.
    Boix L, Rosa JL, Ventura F, et al. c-met mRNA overexpression in human hepatocellular carcinoma. Hepatology. 1994;19:88–91.CrossRefPubMedGoogle Scholar
  11. 11.
    Schena M, Shalon D, Davis RW, Brown PO. Quantitative monitoring of gene expression patterns with a complementary DNA microarray. Science. 1995;270:467–70.CrossRefPubMedGoogle Scholar
  12. 12.
    Lu YJ, Yang J, Noel E, et al. Association between large-scale genomic homozygosity without chromosomal loss and nonseminomatous germ cell tumor development. Cancer Res. 2005;65:9137–41.CrossRefPubMedGoogle Scholar
  13. 13.
    Raghavan M, Lillington DM, Skoulakis S, et al. Genome-wide single nucleotide polymorphism analysis reveals frequent partial uniparental disomy due to somatic recombination in acute myeloid leukemias. Cancer Res. 2005;65:375–8.PubMedGoogle Scholar
  14. 14.
    Kanda M, Nomoto S, Okamura Y, et al. Detection of metallothionein 1G as a methylated tumor suppressor gene in human hepatocellular carcinoma using a novel method of double combination array analysis. Int J Oncol. 2009;35:477–83.CrossRefPubMedGoogle Scholar
  15. 15.
    Okamura Y, Nomoto S, Kanda M, et al. Leukemia inhibitory factor receptor (LIFR) is detected as a novel suppressor gene of hepatocellular carcinoma using double-combination array. Cancer Lett. 2010;28:170–7.CrossRefGoogle Scholar
  16. 16.
    Nomoto S, Kanda M, Okamura Y, et al. Epidermal growth factor–containing fibulin-like extracellular matrix protein 1, EFEMP1, a novel tumor-suppressor gene detected in hepatocellular carcinoma using double combination array analysis. Ann Surg Oncol. 2010;17:923–32.CrossRefPubMedGoogle Scholar
  17. 17.
    Rice DS, Curran T. Role of the reelin signaling pathway in central nervous system development. Annu Rev Neurosci. 2001;24:1005–39.CrossRefPubMedGoogle Scholar
  18. 18.
    Tissir T, Goffnet AM. Reelin and brain development. Nat Rev Neurosci. 2003;4:496–505.CrossRefPubMedGoogle Scholar
  19. 19.
    Sato N, Fukushima N, Chang R, Matsubayashi H, Goggins M. Microarrays and other new technologies. Gastroenterology. 2006;130:548–65.CrossRefPubMedGoogle Scholar
  20. 20.
    Abdolmaleky HM, Cheng KH, Russo A, et al. Hypermethylation of the reelin (RELN) promoter in the brain of schizophrenic patients: a preliminary report. Am J Med Genet B Neuropsychiatr Genet. 2005;134:60–6.Google Scholar
  21. 21.
    Grayson DR, Jia X, Chen Y, et al. Reelin promoter hypermethylation in schizophrenia. Proc Natl Acad Sci USA. 2005;102:9341–6.CrossRefPubMedGoogle Scholar
  22. 22.
    Robertson KD. DNA methylation and chromatin—unraveling the tangled web. Oncogene. 2002;21:5361–79.CrossRefPubMedGoogle Scholar
  23. 23.
    Johnstone RW. Histone-deacetylase inhibitors: novel drugs for the treatment of cancer. Nat Rev Drug Discov. 2002;1:287–99.CrossRefPubMedGoogle Scholar
  24. 24.
    Blaheta RA, Michaelis M, Driever PH, Cinatl J Jr. Evolving anticancer drug valproic acid: insights into the mechanism and clinical studies. Med Res Rev. 2005;25:383–97.CrossRefPubMedGoogle Scholar
  25. 25.
    Kuendgen A, Strupp C, Aivado M, et al. Treatment of myelodysplastic syndromes with valproic acid alone or in combination with all-trans retinoic acid. Blood. 2004;104:1266–9.CrossRefPubMedGoogle Scholar

Copyright information

© Society of Surgical Oncology 2010

Authors and Affiliations

  • Yukiyasu Okamura
    • 1
  • Shuji Nomoto
    • 1
  • Mitsuro Kanda
    • 1
  • Masamichi Hayashi
    • 1
  • Yoko Nishikawa
    • 1
  • Tsutomu Fujii
    • 1
  • Hiroyuki Sugimoto
    • 1
  • Shin Takeda
    • 1
  • Akimasa Nakao
    • 1
  1. 1.Department of Surgery II, Graduate School and MedicineUniversity of NagoyaNagoyaJapan

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