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Role of Stem Cell Factor andc-kitSignaling in Regulation of Fetal Intestinal Epithelial Cell Adhesion to Fibronectin

Authors
Journal
Experimental Cell Research
0014-4827
Publisher
Elsevier
Publication Date
Volume
266
Issue
2
Identifiers
DOI: 10.1006/excr.2001.5221
Keywords
  • Stem Cell Factor
  • C-Kit
  • Fibronectin
  • Vla-5
  • Intestinal Epithelial Cell
  • Mesenchymes
Disciplines
  • Design

Abstract

Abstract The interaction of stem cell factor (SCF) and c-kit is considered to be an important signaling event for the homeostasis of the epithelial barrier function in the intestinal tract. This study was designed to investigate the role of the SCF and c-kit signaling pathway in adhesion of intestinal epithelial cells (IECs) to fibronectin (FN) using primary cells. Fetal murine IECs were prepared from the small intestine of mouse fetus. The mRNAs coding for SCF in mesenchymes and c-kit in IECs were detected by reverse transcription-PCR. The expression of FN receptor VLA-5 on IECs was examined by flow cytometry. A cell adhesion assay showed that the stimulation of IECs with SCF increased the number of cells adhering to FN. Experiments using specific antibody against SCF indicated that this increase in cell adhesion was SCF-dependent. On the other hand, SCF did not influence the expression of VLA-5 on IECs. The IEC adhesion to FN was inhibited by specific antibody against the FN receptor (VLA-5), as well as competitive Arg-Gly-Asp (RGD) peptide. When alteration of intracellular signal transduction induced by SCF was examined, it was found that SCF stimulated a tyrosine-specific c-kit autophosphorylation cascade of IECs. Further, preincubation of IECs with an optimal concentration of genistein resulted in the inhibition of SCF-induced c-kit phosphorylation and adhesion of IECs to FN. These results suggested that adhesion of immature IECs to FN is regulated by activation of RGD-dependent VLA-5 through the SCF and c-kit signal transduction pathway. SCF, which may be produced by mesenchymes locally, is an important regulatory factor for the adhesion of immature IECs to basement membrane matrix via VLA-5 and FN interaction. This cytokine-regulated interaction between VLA-5 and FN may play an important role in the development and wound repair of the intestinal tract.

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