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  • Oncogenomics
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Frequent methylation-associated silencing of a candidate tumor-suppressor, CRABP1, in esophageal squamous-cell carcinoma

Oncogene volume 26, pages 6456–6468 (2007)Cite this article

Abstract

Epigenetic alterations and the resulting inactivation of tumor suppressor genes often contribute to the development of various cancers. To identify novel candidates that may be silenced by aberrant methylation in esophageal squamous-cell carcinoma (ESCC), we analysed ESCC cell lines by a recently developed method known as bacterial artificial chromosome array-based methylated CpG island amplification (BAMCA), and selected candidates through BAMCA-assisted strategy. In the course of this program, we identified frequent CpG methylation-dependent silencing of the gene encoding cellular retinoic acid binding protein 1 (CRABP1) in our panel of ESCC cell lines. Expression of CRABP1 mRNA was restored in gene-silenced ESCC cells after treatment with 5-aza 2′-deoxycytidine. The DNA methylation status of the CRABP1 CpG island with clear promoter activity correlated inversely with expression of this gene. CpG methylation of CRABP1 was frequently observed in primary ESCC tissues as well. Restoration of CRABP1 expression in ESCC cells lacking the protein reduced cell growth by inducing arrest at G0–G1, whereas knockdown of the gene in cells expressing CRABP1 promoted cell growth. Among 113 primary ESCC tumors, the absence of immunoreactive CRABP1 was significantly associated with de-differentiation of cancer cells and with distant lymph-node metastases in the patients. These results indicate that CRABP1 appears to have a tumor-suppressor function in esophageal epithelium, and its epigenetic silencing may play a pivotal role during esophageal carcinogenesis. Its expression status in biopsies or resected tumors might serve as an index for identifying ESCC patients for whom combined therapeutic modalities would be recommended.

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Acknowledgements

This work is supported by Grants-in-aid for scientific research on priority areas and 21st Century center of excellence program for molecular destruction and reconstitution of tooth and bone from the Ministry of Education, Culture, Sports, Science, and Technology, Japan and a grant-in-aid from core research for evolutional science and technology (CREST) of the Japan science and technology corporation (JST).

We are grateful to Professor Yusuke Nakamura (Human Genome Center, Institute of Medical Science, The University of Tokyo) for continuous encouragement throughout this work. We also thank Ayako Takahashi, and Rumi Mori for technical assistance, and Yoriko Fukukawa for secretarial assistance.

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Authors and Affiliations

  1. Department of Molecular Cytogenetics, Medical Research Institute and School of Biomedical Science, Tokyo Medical and Dental University, Tokyo, Japan

    K Tanaka, I Imoto, J Inoue, K Kozaki & J Inazawa

  2. Department of Vascular and Applied Surgery, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan

    K Tanaka, T Iwai & T Kawano

  3. Department of Genome Medicine, Hard Tissue Genome Research Center, Tokyo Medical and Dental University, Tokyo, Japan

    I Imoto, K Kozaki & J Inazawa

  4. Core Research for Evolutional Science and Technology (CREST) of Japan Science and Technology Corporation (JST), Kawaguchi, Japan

    I Imoto, J Inoue, H Tsuda & J Inazawa

  5. Department of Basic Pathology, National Defense Medical College, Tokorozawa, Japan

    H Tsuda

  6. First Department of Surgery, Hyogo College of Medicine, Nishinomiya, Japan

    Y Shimada

  7. Department of Surgery, National Defense Medical College, Tokorozawa, Japan

    S Aiko & Y Yoshizumi

  8. 21st Century Center of Excellence Program for Molecular Destruction and Reconstitution of Tooth and Bone, Tokyo Medical and Dental University, Tokyo, Japan

    J Inazawa

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Correspondence to J Inazawa.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Tanaka, K., Imoto, I., Inoue, J. et al. Frequent methylation-associated silencing of a candidate tumor-suppressor, CRABP1, in esophageal squamous-cell carcinoma. Oncogene 26, 6456–6468 (2007). https://doi.org/10.1038/sj.onc.1210459

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