Regulation of human Cripto-1 gene expression by TGF-beta1 and BMP-4 in embryonal and colon cancer cells

J Cell Physiol. 2008 Apr;215(1):192-203. doi: 10.1002/jcp.21301.


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-beta family members, TGF-beta1 and BMP-4. In particular, TGF-beta1 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-beta1 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-beta1 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-beta family.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5' Flanking Region
  • Animals
  • Base Sequence
  • Bone Morphogenetic Protein 4
  • Bone Morphogenetic Proteins / pharmacology*
  • Cell Movement / drug effects
  • Cell Proliferation / drug effects
  • Colonic Neoplasms / genetics*
  • Colonic Neoplasms / pathology
  • Embryonal Carcinoma Stem Cells / metabolism*
  • Embryonal Carcinoma Stem Cells / pathology
  • Epidermal Growth Factor / deficiency
  • Epidermal Growth Factor / genetics*
  • Epidermal Growth Factor / metabolism
  • GPI-Linked Proteins
  • Gene Expression Regulation, Neoplastic / drug effects*
  • Humans
  • Intercellular Signaling Peptides and Proteins
  • Membrane Glycoproteins / deficiency
  • Membrane Glycoproteins / genetics*
  • Membrane Glycoproteins / metabolism
  • Mice
  • Molecular Sequence Data
  • Mutation / genetics
  • Neoplasm Proteins / deficiency
  • Neoplasm Proteins / genetics*
  • Neoplasm Proteins / metabolism
  • Promoter Regions, Genetic
  • Protein Binding / drug effects
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Smad Proteins / metabolism
  • Transforming Growth Factor beta1 / pharmacology*


  • BMP4 protein, human
  • Bmp4 protein, mouse
  • Bone Morphogenetic Protein 4
  • Bone Morphogenetic Proteins
  • GPI-Linked Proteins
  • Intercellular Signaling Peptides and Proteins
  • Membrane Glycoproteins
  • Neoplasm Proteins
  • RNA, Messenger
  • Smad Proteins
  • TDGF1 protein, human
  • Tdgf1 protein, mouse
  • Transforming Growth Factor beta1
  • Epidermal Growth Factor