TY - JOUR
T1 - Cdx2 and the Brm-type SWI/SNF complex cooperatively regulate villin expression in gastrointestinal cells
AU - Yamamichi, Nobutake
AU - Inada, Ken ichi
AU - Furukawa, Chihiro
AU - Sakurai, Kohei
AU - Tando, Toshio
AU - Ishizaka, Aya
AU - Haraguchi, Takeshi
AU - Mizutani, Taketoshi
AU - Fujishiro, Mitsuhiro
AU - Shimomura, Ryoichi
AU - Oka, Masashi
AU - Ichinose, Masao
AU - Tsutsumi, Yutaka
AU - Omata, Masao
AU - Iba, Hideo
N1 - Funding Information:
Grant support/financial disclosures: This work was supported in part by a Grant-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Cultures, Sports, Science, and Technology of Japan; and was also supported in part by Strategic cooperation to control emerging and reemerging infections funded by the Special Coordination Funds for Promoting Science and Technology of Ministry of Education, Culture, Sports, Science and Technology (MEXT). None of the authors have any conflicts of interest associated with this study. We disclosed potential conflicts to all the study participants.
PY - 2009/6/10
Y1 - 2009/6/10
N2 - In our recent study showing a correlation between Brm-deficiency and undifferentiated status of gastric cancer, we found that the Brm-type SWI/SNF complex is required for villin expression. To elucidate intestinal villin regulation more precisely, we here analyzed structure and function of the promoter of human villin. About 1.1 kb upstream of the determined major transcription start site, we identified a highly conserved region (HCR-Cdx) among mammals, which contains two binding sites for Cdx. Expression analyses of 30 human gastrointestinal cell lines suggested that villin is regulated by Cdx2. Introduction of Cdx family genes into colorectal SW480 cells revealed that villin is strongly induced strongly by Cdx2, moderately by Cdx1, and marginally by Cdx4. Knockdown of Cdx2 in SW480 cells caused a clear downregulation of villin, and reporter assays showed that HCR-Cdx is crucial for Cdx2-dependent and Brm-dependent villin expression. Immunohistochemical analyses of gastric intestinal metaplasia and cancer revealed that villin and Cdx2 expression are tightly coupled. GST pull-down assays demonstrated a direct interaction between Cdx2 and several SWI/SNF subunits. Chromatin immunoprecipitation analyses showed the recruitment of Cdx2 and Brm around HCR-Cdx. From these results, we concluded that Cdx2 regulates intestinal villin expression through recruiting Brm-type SWI/SNF complex to the villin promoter.
AB - In our recent study showing a correlation between Brm-deficiency and undifferentiated status of gastric cancer, we found that the Brm-type SWI/SNF complex is required for villin expression. To elucidate intestinal villin regulation more precisely, we here analyzed structure and function of the promoter of human villin. About 1.1 kb upstream of the determined major transcription start site, we identified a highly conserved region (HCR-Cdx) among mammals, which contains two binding sites for Cdx. Expression analyses of 30 human gastrointestinal cell lines suggested that villin is regulated by Cdx2. Introduction of Cdx family genes into colorectal SW480 cells revealed that villin is strongly induced strongly by Cdx2, moderately by Cdx1, and marginally by Cdx4. Knockdown of Cdx2 in SW480 cells caused a clear downregulation of villin, and reporter assays showed that HCR-Cdx is crucial for Cdx2-dependent and Brm-dependent villin expression. Immunohistochemical analyses of gastric intestinal metaplasia and cancer revealed that villin and Cdx2 expression are tightly coupled. GST pull-down assays demonstrated a direct interaction between Cdx2 and several SWI/SNF subunits. Chromatin immunoprecipitation analyses showed the recruitment of Cdx2 and Brm around HCR-Cdx. From these results, we concluded that Cdx2 regulates intestinal villin expression through recruiting Brm-type SWI/SNF complex to the villin promoter.
UR - https://www.scopus.com/pages/publications/67349226702
UR - https://www.scopus.com/inward/citedby.url?scp=67349226702&partnerID=8YFLogxK
U2 - 10.1016/j.yexcr.2009.01.006
DO - 10.1016/j.yexcr.2009.01.006
M3 - Article
C2 - 19371634
AN - SCOPUS:67349226702
SN - 0014-4827
VL - 315
SP - 1779
EP - 1789
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 10
ER -
