Cooperation between HNF-1alpha, Cdx2, and GATA-4 in initiating an enterocytic differentiation program in a normal human intestinal epithelial progenitor cell line

Am J Physiol Gastrointest Liver Physiol. 2010 Apr;298(4):G504-17. doi: 10.1152/ajpgi.00265.2009. Epub 2010 Feb 4.

Abstract

In the intestinal epithelium, the Cdx, GATA, and HNF transcription factor families are responsible for the expression of differentiation markers such as sucrase-isomaltase. Although previous studies have shown that Cdx2 can induce differentiation in rat intestinal IEC-6 cells, no data are available concerning the direct implication of transcription factors on differentiation in human normal intestinal epithelial cell types. We investigated the role of Cdx2, GATA-4, and HNF-1alpha using the undifferentiated human intestinal epithelial crypt cell line HIEC. These transcription factors were tested on proliferation and expression of polarization and differentiation markers. Ectopic expression of Cdx2 or HNF-1alpha, alone or in combination, altered cell proliferation abilities through the regulation of cyclin D1 and p27 expression. HNF-1alpha and GATA-4 together induced morphological modifications of the cells toward polarization, resulting in the appearance of functional features such as microvilli. HNF-1alpha was also sufficient to induce the expression of cadherins and dipeptidylpeptidase, whereas in combination with Cdx2 it allowed the expression of the late differentiation marker sucrase-isomaltase. Large-scale analysis of gene expression confirmed the cooperative effect of these factors. Finally, although DcamKL1 and Musashi-1 expression were downregulated in differentiated HIEC, other intestinal stem cell markers, such as Bmi1, were unaffected. These observations show that, in cooperation with Cdx2, HNF-1alpha acts as a key factor on human intestinal cells to trigger the onset of their functional differentiation program whereas GATA-4 appears to promote morphological changes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CDX2 Transcription Factor
  • Cadherins / genetics
  • Cadherins / metabolism
  • Cell Differentiation / physiology*
  • Cell Line
  • Cell Proliferation
  • Cyclin D1 / metabolism
  • Cyclin-Dependent Kinase Inhibitor p27
  • Dipeptidyl Peptidase 4 / genetics
  • Down-Regulation / genetics
  • Enterocytes / cytology*
  • Enterocytes / metabolism
  • Enterocytes / ultrastructure
  • GATA4 Transcription Factor / genetics
  • GATA4 Transcription Factor / metabolism*
  • Gene Expression / genetics
  • Gene Expression Profiling
  • Hepatocyte Nuclear Factor 1-alpha / genetics
  • Hepatocyte Nuclear Factor 1-alpha / metabolism*
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism*
  • Humans
  • Interphase / genetics
  • Intestinal Mucosa / cytology*
  • Intestinal Mucosa / metabolism
  • Intestine, Small / cytology
  • Intestine, Small / metabolism
  • Intracellular Signaling Peptides and Proteins / genetics
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Nerve Tissue Proteins / genetics
  • Nuclear Proteins / genetics
  • Polycomb Repressive Complex 1
  • Protein-Serine-Threonine Kinases / genetics
  • Proto-Oncogene Proteins / genetics
  • RNA-Binding Proteins / genetics
  • Receptors, G-Protein-Coupled / genetics
  • Repressor Proteins / genetics
  • Stem Cells / cytology*
  • Stem Cells / metabolism
  • Stem Cells / ultrastructure
  • Sucrase-Isomaltase Complex / genetics
  • Transfection
  • Up-Regulation / genetics

Substances

  • BMI1 protein, human
  • CCND1 protein, human
  • CDH17 protein, human
  • CDKN1B protein, human
  • CDX2 Transcription Factor
  • CDX2 protein, human
  • Cadherins
  • GATA4 Transcription Factor
  • HNF1A protein, human
  • Hepatocyte Nuclear Factor 1-alpha
  • Homeodomain Proteins
  • Intracellular Signaling Peptides and Proteins
  • LGR5 protein, human
  • MSI1 protein, human
  • Nerve Tissue Proteins
  • Nuclear Proteins
  • Proto-Oncogene Proteins
  • RNA-Binding Proteins
  • Receptors, G-Protein-Coupled
  • Repressor Proteins
  • Cyclin D1
  • Cyclin-Dependent Kinase Inhibitor p27
  • Polycomb Repressive Complex 1
  • DCLK1 protein, human
  • Protein-Serine-Threonine Kinases
  • Sucrase-Isomaltase Complex
  • DPP4 protein, human
  • Dipeptidyl Peptidase 4