Regulation of human cripto-1 gene expression by TGF-β1 and BMP-4 in embryonal and colon cancer cells

Human Cripto‐1 (CR‐1) is a cell membrane protein that is overexpressed in several different types of human carcinomas. In the present study we investigated the mechanisms that regulate the expression of CR‐1 gene in cancer cells. We cloned a 2,481 bp 5′‐flanking region of the human CR‐1 gene into a...

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Published in:Journal of cellular physiology 2008-04, Vol.215 (1), p.192-203
Main Authors: Mancino, Mario, Strizzi, Luigi, Wechselberger, Christian, Watanabe, Kazuhide, Gonzales, Monica, Hamada, Shin, Normanno, Nicola, Salomon, David S., Bianco, Caterina
Format: Article
Language:English
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Summary:Human Cripto‐1 (CR‐1) is a cell membrane protein that is overexpressed in several different types of human carcinomas. In the present study we investigated the mechanisms that regulate the expression of CR‐1 gene in cancer cells. We cloned a 2,481 bp 5′‐flanking region of the human CR‐1 gene into a luciferase reporter vector and transfected NTERA‐2 human embryonal carcinoma cells and LS174‐T colon cancer cells to test for promoter activity. Activity of CR‐1 promoter in both cell lines was modulated by two TGF‐β family members, TGF‐β1 and BMP‐4. In particular, TGF‐β1 significantly up‐regulated CR‐1 promoter activity, whereas a dramatic reduction in CR‐1 promoter activity was observed with BMP‐4 in NTERA‐2 and LS174‐T cells. Changes in the CR‐1 promoter activity following TGF‐β1 and BMP‐4 treatments correlated with changes in CR‐1 mRNA and protein expression in NTERA‐2 and LS174‐T cells. We also identified three Smad binding elements (SBEs) within the CR‐1 promoter and point mutation of SBE1 (−2,197/−2,189) significantly reduced response of the CR‐1 promoter to both TGF‐β1 and BMP‐4 in NTERA‐2 and LS174‐T cells. Chromatin immunoprecipitation assay also demonstrated binding of Smad‐4 to a CR‐1 promoter DNA sequence containing SBE1 in LS174‐T cells. Finally, BMP‐4 inhibited migration of LS174‐T cells and F9 mouse embryonal carcinoma cells by downregulation of CR‐1 protein. In conclusion, these results suggest a differential modulation of CR‐1 gene expression in embryonal and colon cancer cells by two different members of the TGF‐β family. J. Cell. Physiol. 215: 192–203, 2008. © 2007 Wiley‐Liss, Inc.
ISSN:0021-9541
1097-4652
DOI:10.1002/jcp.21301