Aberrant promoter methylation in human DAB2 interactive protein (hDAB2IP) gene in gastrointestinal tumour

H. Dote, Shinichi Toyooka, K. Tsukuda, M. Yano, T. Ota, M. Murakami, M. Naito, M. Toyota, A. F. Gazdar, N. Shimizu

Research output: Contribution to journalArticle

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Abstract

The human DOC-2/DAB2 interactive protein (hDAB2IP) gene is a novel member of the Ras GTPase-activating family and has been demonstrated to be a tumour-suppressor gene inactivated by methylation in several cancers. In this study, we analysed the methylation and expression status of hDAB2IP in gastrointestinal tumours. The promoter region of hDAB2IP was divided into two regions (m2a and m2b) based on our previous report, and the methylation status was determined by bisulphite DNA sequencing in gastric cancer cell lines. The gene expression was semiquantified by real-time RT-PCR, and the results indicated that the m2b promoter region might be an authentic methylation-mediated key regulator of the gene expression. Based on the sequence data, we developed a methylation-specific PCR (MSP) for the m2a and m2b regions and applied it to the samples. Methylation-specific PCR revealed aberrant methylation in the m2a region in eight of 12 gastric cancer cell lines (67%), 16 of 35 gastric cancer tissues (46%) and 29 of 60 colorectal cancer tissues (48%), and in the m2b region in eight of 12 cell lines (67%), I 5 of 35 gastric cancer tissues (43%) and 28 of 60 colorectal cancer tissues (47%). On the other hand, seven (12%) and I I (19%) of 59 gastrointestinal nonmalignant mucosal specimens showed methylation in the m2a and m2b regions, respectively, suggesting that hDAB2IP methylation might play a causative role in carcinogenesis. The 5-aza-2′-deoxycytidine treatment restored the gene expression in the m2b-methylated cell lines, confirming that the methylation caused gene downregulation. We also examined the relationship between hDAB2IP methylation and the clinicopathological features in patients with primary tumours, and determined that methylation in the m2b region was associated with location of the tumour in the stomach. In summary, our results demonstrated that hDAB2IP methylation is frequently present in gastrointestinal tumours and that the resulting gene silencing plays an important role in gastrointestinal carcinogenesis.

Original languageEnglish
Pages (from-to)1117-1125
Number of pages9
JournalBritish Journal of Cancer
Volume92
Issue number6
DOIs
Publication statusPublished - Mar 28 2005

Fingerprint

Methylation
Genes
Neoplasms
Stomach Neoplasms
Cell Line
decitabine
Gene Expression
Genetic Promoter Regions
human DAB2 protein
Colorectal Neoplasms
Carcinogenesis
ras Proteins
Polymerase Chain Reaction
Gene Silencing
Regulator Genes
Tumor Suppressor Genes
DNA Sequence Analysis
Real-Time Polymerase Chain Reaction
Stomach
Down-Regulation

Keywords

  • Gastrointestinal tumour
  • hDAB2IP
  • Histone acetylation
  • Methylation
  • Methylation specific PCR

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Aberrant promoter methylation in human DAB2 interactive protein (hDAB2IP) gene in gastrointestinal tumour. / Dote, H.; Toyooka, Shinichi; Tsukuda, K.; Yano, M.; Ota, T.; Murakami, M.; Naito, M.; Toyota, M.; Gazdar, A. F.; Shimizu, N.

In: British Journal of Cancer, Vol. 92, No. 6, 28.03.2005, p. 1117-1125.

Research output: Contribution to journalArticle

Dote, H, Toyooka, S, Tsukuda, K, Yano, M, Ota, T, Murakami, M, Naito, M, Toyota, M, Gazdar, AF & Shimizu, N 2005, 'Aberrant promoter methylation in human DAB2 interactive protein (hDAB2IP) gene in gastrointestinal tumour', British Journal of Cancer, vol. 92, no. 6, pp. 1117-1125. https://doi.org/10.1038/sj.bjc.6602458
Dote, H. ; Toyooka, Shinichi ; Tsukuda, K. ; Yano, M. ; Ota, T. ; Murakami, M. ; Naito, M. ; Toyota, M. ; Gazdar, A. F. ; Shimizu, N. / Aberrant promoter methylation in human DAB2 interactive protein (hDAB2IP) gene in gastrointestinal tumour. In: British Journal of Cancer. 2005 ; Vol. 92, No. 6. pp. 1117-1125.
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abstract = "The human DOC-2/DAB2 interactive protein (hDAB2IP) gene is a novel member of the Ras GTPase-activating family and has been demonstrated to be a tumour-suppressor gene inactivated by methylation in several cancers. In this study, we analysed the methylation and expression status of hDAB2IP in gastrointestinal tumours. The promoter region of hDAB2IP was divided into two regions (m2a and m2b) based on our previous report, and the methylation status was determined by bisulphite DNA sequencing in gastric cancer cell lines. The gene expression was semiquantified by real-time RT-PCR, and the results indicated that the m2b promoter region might be an authentic methylation-mediated key regulator of the gene expression. Based on the sequence data, we developed a methylation-specific PCR (MSP) for the m2a and m2b regions and applied it to the samples. Methylation-specific PCR revealed aberrant methylation in the m2a region in eight of 12 gastric cancer cell lines (67{\%}), 16 of 35 gastric cancer tissues (46{\%}) and 29 of 60 colorectal cancer tissues (48{\%}), and in the m2b region in eight of 12 cell lines (67{\%}), I 5 of 35 gastric cancer tissues (43{\%}) and 28 of 60 colorectal cancer tissues (47{\%}). On the other hand, seven (12{\%}) and I I (19{\%}) of 59 gastrointestinal nonmalignant mucosal specimens showed methylation in the m2a and m2b regions, respectively, suggesting that hDAB2IP methylation might play a causative role in carcinogenesis. The 5-aza-2′-deoxycytidine treatment restored the gene expression in the m2b-methylated cell lines, confirming that the methylation caused gene downregulation. We also examined the relationship between hDAB2IP methylation and the clinicopathological features in patients with primary tumours, and determined that methylation in the m2b region was associated with location of the tumour in the stomach. In summary, our results demonstrated that hDAB2IP methylation is frequently present in gastrointestinal tumours and that the resulting gene silencing plays an important role in gastrointestinal carcinogenesis.",
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AU - Ota, T.

AU - Murakami, M.

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N2 - The human DOC-2/DAB2 interactive protein (hDAB2IP) gene is a novel member of the Ras GTPase-activating family and has been demonstrated to be a tumour-suppressor gene inactivated by methylation in several cancers. In this study, we analysed the methylation and expression status of hDAB2IP in gastrointestinal tumours. The promoter region of hDAB2IP was divided into two regions (m2a and m2b) based on our previous report, and the methylation status was determined by bisulphite DNA sequencing in gastric cancer cell lines. The gene expression was semiquantified by real-time RT-PCR, and the results indicated that the m2b promoter region might be an authentic methylation-mediated key regulator of the gene expression. Based on the sequence data, we developed a methylation-specific PCR (MSP) for the m2a and m2b regions and applied it to the samples. Methylation-specific PCR revealed aberrant methylation in the m2a region in eight of 12 gastric cancer cell lines (67%), 16 of 35 gastric cancer tissues (46%) and 29 of 60 colorectal cancer tissues (48%), and in the m2b region in eight of 12 cell lines (67%), I 5 of 35 gastric cancer tissues (43%) and 28 of 60 colorectal cancer tissues (47%). On the other hand, seven (12%) and I I (19%) of 59 gastrointestinal nonmalignant mucosal specimens showed methylation in the m2a and m2b regions, respectively, suggesting that hDAB2IP methylation might play a causative role in carcinogenesis. The 5-aza-2′-deoxycytidine treatment restored the gene expression in the m2b-methylated cell lines, confirming that the methylation caused gene downregulation. We also examined the relationship between hDAB2IP methylation and the clinicopathological features in patients with primary tumours, and determined that methylation in the m2b region was associated with location of the tumour in the stomach. In summary, our results demonstrated that hDAB2IP methylation is frequently present in gastrointestinal tumours and that the resulting gene silencing plays an important role in gastrointestinal carcinogenesis.

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