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Aberrant promoter methylation and tumor suppressive activity of the DFNA5 gene in colorectal carcinoma

Oncogene volume 27, pages 3624–3634 (2008)Cite this article

Abstract

To identify novel methylated gene promoters, we compared differential RNA expression profiles of colorectal cancer (CRC) cell lines with or without treatment of 5-aza-2′-deoxycytidine (5-aza-dC). Out of 1776 genes that were initially ‘absent (that is, silenced)’ by gene expression array analysis, we selected 163 genes that were increased after 5-aza-dC treatment in at least two of three CRC cell lines. The microarray results were confirmed by Reverse Transcription–PCR, and CpG island of the gene promoters were amplified and sequenced for examination of cancer-specific methylation. Among the genes identified, the deafness, autosomal dominant 5 gene, DFNA5, promoter was found to be methylated in primary tumor tissues with high frequency (65%, 65/100). Quantitative methylation-specific PCR of DFNA5 clearly discriminated primary CRC tissues from normal colon tissues (3%, 3/100). The mRNA expression of DFNA5 in four of five colon cancer tissues was significantly downregulated as compared to normal tissues. Moreover, forced expression of full-length DFNA5 in CRC cell lines markedly decreased the cell growth and colony-forming ability whereas knockdown of DFNA5 increased cell growth in culture. Our data implicate DFNA5 as a novel tumor suppressor gene in CRC and a valuable molecular marker for human cancer.

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Acknowledgements

Under a licensing agreement between OncoMethylome Sciences, SA and the Johns Hopkins University, Dr Sidransky is entitled to a share of royalty received by the university on sales of products described in this article. Dr Sidransky owns OncoMethylome Sciences, SA stock, which is subject to certain restrictions under the university policy. Dr Sidransky is a paid consultant to OncoMethylome Sciences, SA and is a paid member of the company's Scientific Advisory Board. The term of this arrangement is being managed by the Johns Hopkins University in accordance with its conflict of interest policies.

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Authors and Affiliations

  1. Department of Otolaryngology, Head and Neck Cancer Research Division, Johns Hopkins University, Baltimore, MD, USA

    M S Kim, X Chang, K Yamashita, J K Nagpal, B Trink, E A Ratovitski & D Sidransky

  2. Department of Genetic Medicine, Institute of Cell Engineering, Johns Hopkins University, Baltimore, MD, USA

    J H Baek

  3. Department of Pathology, Karmanos Cancer Institute, Wayne State University, Detroit, MI, USA

    G Wu

  4. Department of Surgical Oncology, Medical Institute of Bioregulation, Kyushu University, Tsurumibaru, Beppu, Japan

    M Mori

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  1. M S Kim
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  2. X Chang
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  3. K Yamashita
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  4. J K Nagpal
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  5. J H Baek
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  6. G Wu
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  7. B Trink
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  8. E A Ratovitski
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  10. D Sidransky
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Correspondence to D Sidransky.

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

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Kim, M., Chang, X., Yamashita, K. et al. Aberrant promoter methylation and tumor suppressive activity of the DFNA5 gene in colorectal carcinoma. Oncogene 27, 3624–3634 (2008). https://doi.org/10.1038/sj.onc.1211021

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