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.
PURPOSE:HumanDOC-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.
Authors: Sara Koenig McLaughlin; Sarah Naomi Olsen; Benjamin Dake; Thomas De Raedt; Elgene Lim; Roderick Terry Bronson; Rameen Beroukhim; Kornelia Polyak; Myles Brown; Charlotte Kuperwasser; Karen Cichowski Journal: Cancer Cell Date: 2013-09-09 Impact factor: 31.743
Authors: Kiyoshi Misawa; Yo Ueda; Takeharu Kanazawa; Yuki Misawa; Ilwhan Jang; John Chadwick Brenner; Tetsuya Ogawa; Satoru Takebayashi; Reidar A Grenman; James G Herman; Hiroyuki Mineta; Thomas E Carey Journal: Clin Cancer Res Date: 2008-12-01 Impact factor: 12.531
Authors: J Zhou; J Luo; K Wu; E-J Yun; P Kapur; R-C Pong; Y Du; B Wang; C Authement; E Hernandez; J Yang; G Xiao; T-L Cha; H-C Wu; D Wu; V Margulis; Y Lotan; J Brugarolas; D He; J-T Hsieh Journal: Oncogene Date: 2016-02-15 Impact factor: 9.867
Authors: Daxing Xie; Crystal Gore; Jun Liu; Rey-Chen Pong; Ralph Mason; Guiyang Hao; Michael Long; Wareef Kabbani; Luyang Yu; Haifeng Zhang; Hong Chen; Xiankai Sun; David A Boothman; Wang Min; Jer-Tsong Hsieh Journal: Proc Natl Acad Sci U S A Date: 2010-01-13 Impact factor: 11.205