Aberrant Promoter Methylation in Human DAB2 Interactive Protein (hDAB2IP) Gene in Breast Cancer

Hideaki Dote, Shinichi Toyooka, Kazunori Tsukuda, Masaaki Yano, Mamoru Oouchida, Hiroyoshi Doihara, Makoto Suzuki, Hong Chen, Jer Tsong Hsieh, Adi F. Gazdar, Nobuyoshi Shimizu

Research output: Contribution to journalArticle

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Abstract

Purpose: 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 tumor suppressor gene inactivated by methylation in prostate cancer. We analyzed methylation and expression status of hDAB2IP in breast cancer. Experimental Design: The promoter region of hDAB2IP was divided into two regions (m2a and m2b) following our previous report on prostate cancer, and methylation status was determined in breast cancer cell lines with bisulfited DNA sequencing. Expression was semiquantified with real-time reverse transcription-PCR to find that aberrant methylation showed the inverse relationship with expression. On the basis of sequence data, we developed methylation-specific PCR for m2a and m2b regions and applied to samples. Results: Aberrant methylation was detected in 11 of 25 breast cancer cell lines (44%) and 15 of 39 primary tumors (38%) at the m2a region and in 12 of 25 cell lines (48%) and 13 of 39 tumors (33%) at the m2b region. In addition, gene expression was restored in methylated cell lines with 5-aza-2′ -deoxycytidine, confirming that methylation caused gene down-regulation. We also examined the relationship between hDAB2IP methylation and clinicopathologic features in primary tumors and found that methylation in the m2b region was associated with progressive nodal status of tumors. Conclusions: We developed methylation-specific PCR for hDAB2IP and examined its methylation status in breast cancer. Our results demonstrate that hDAB2IP methylation frequently is present in breast cancer and plays a key role in hDAB2IP inactivation, suggesting the relationship between hDAB2IP methylation and lymph node metastasis of breast cancer.

Original languageEnglish
Pages (from-to)2082-2089
Number of pages8
JournalClinical Cancer Research
Volume10
Issue number6
DOIs
Publication statusPublished - Mar 15 2004

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Methylation
Breast Neoplasms
Genes
Cell Line
decitabine
human DAB2 protein
Polymerase Chain Reaction
Neoplasms
Prostatic Neoplasms
ras Proteins
Tumor Suppressor Genes
DNA Sequence Analysis
Genetic Promoter Regions
Reverse Transcription
Research Design
Down-Regulation
Lymph Nodes
Neoplasm Metastasis
Gene Expression

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Aberrant Promoter Methylation in Human DAB2 Interactive Protein (hDAB2IP) Gene in Breast Cancer. / Dote, Hideaki; Toyooka, Shinichi; Tsukuda, Kazunori; Yano, Masaaki; Oouchida, Mamoru; Doihara, Hiroyoshi; Suzuki, Makoto; Chen, Hong; Hsieh, Jer Tsong; Gazdar, Adi F.; Shimizu, Nobuyoshi.

In: Clinical Cancer Research, Vol. 10, No. 6, 15.03.2004, p. 2082-2089.

Research output: Contribution to journalArticle

Dote, Hideaki ; Toyooka, Shinichi ; Tsukuda, Kazunori ; Yano, Masaaki ; Oouchida, Mamoru ; Doihara, Hiroyoshi ; Suzuki, Makoto ; Chen, Hong ; Hsieh, Jer Tsong ; Gazdar, Adi F. ; Shimizu, Nobuyoshi. / Aberrant Promoter Methylation in Human DAB2 Interactive Protein (hDAB2IP) Gene in Breast Cancer. In: Clinical Cancer Research. 2004 ; Vol. 10, No. 6. pp. 2082-2089.
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T1 - Aberrant Promoter Methylation in Human DAB2 Interactive Protein (hDAB2IP) Gene in Breast Cancer

AU - Dote, Hideaki

AU - Toyooka, Shinichi

AU - Tsukuda, Kazunori

AU - Yano, Masaaki

AU - Oouchida, Mamoru

AU - Doihara, Hiroyoshi

AU - Suzuki, Makoto

AU - Chen, Hong

AU - Hsieh, Jer Tsong

AU - Gazdar, Adi F.

AU - Shimizu, Nobuyoshi

PY - 2004/3/15

Y1 - 2004/3/15

N2 - Purpose: 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 tumor suppressor gene inactivated by methylation in prostate cancer. We analyzed methylation and expression status of hDAB2IP in breast cancer. Experimental Design: The promoter region of hDAB2IP was divided into two regions (m2a and m2b) following our previous report on prostate cancer, and methylation status was determined in breast cancer cell lines with bisulfited DNA sequencing. Expression was semiquantified with real-time reverse transcription-PCR to find that aberrant methylation showed the inverse relationship with expression. On the basis of sequence data, we developed methylation-specific PCR for m2a and m2b regions and applied to samples. Results: Aberrant methylation was detected in 11 of 25 breast cancer cell lines (44%) and 15 of 39 primary tumors (38%) at the m2a region and in 12 of 25 cell lines (48%) and 13 of 39 tumors (33%) at the m2b region. In addition, gene expression was restored in methylated cell lines with 5-aza-2′ -deoxycytidine, confirming that methylation caused gene down-regulation. We also examined the relationship between hDAB2IP methylation and clinicopathologic features in primary tumors and found that methylation in the m2b region was associated with progressive nodal status of tumors. Conclusions: We developed methylation-specific PCR for hDAB2IP and examined its methylation status in breast cancer. Our results demonstrate that hDAB2IP methylation frequently is present in breast cancer and plays a key role in hDAB2IP inactivation, suggesting the relationship between hDAB2IP methylation and lymph node metastasis of breast cancer.

AB - Purpose: 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 tumor suppressor gene inactivated by methylation in prostate cancer. We analyzed methylation and expression status of hDAB2IP in breast cancer. Experimental Design: The promoter region of hDAB2IP was divided into two regions (m2a and m2b) following our previous report on prostate cancer, and methylation status was determined in breast cancer cell lines with bisulfited DNA sequencing. Expression was semiquantified with real-time reverse transcription-PCR to find that aberrant methylation showed the inverse relationship with expression. On the basis of sequence data, we developed methylation-specific PCR for m2a and m2b regions and applied to samples. Results: Aberrant methylation was detected in 11 of 25 breast cancer cell lines (44%) and 15 of 39 primary tumors (38%) at the m2a region and in 12 of 25 cell lines (48%) and 13 of 39 tumors (33%) at the m2b region. In addition, gene expression was restored in methylated cell lines with 5-aza-2′ -deoxycytidine, confirming that methylation caused gene down-regulation. We also examined the relationship between hDAB2IP methylation and clinicopathologic features in primary tumors and found that methylation in the m2b region was associated with progressive nodal status of tumors. Conclusions: We developed methylation-specific PCR for hDAB2IP and examined its methylation status in breast cancer. Our results demonstrate that hDAB2IP methylation frequently is present in breast cancer and plays a key role in hDAB2IP inactivation, suggesting the relationship between hDAB2IP methylation and lymph node metastasis of breast cancer.

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