Effect of Tyrosine Kinase Inhibition on Basal and Epidermal Growth Factor-Stimulated Human Caco-2 Enterocyte Sheet Migration and Proliferation

J Cell Physiol. 1994 Sep;160(3):491-501. doi: 10.1002/jcp.1041600312.

Abstract

Mucosal healing requires enterocyte migration (restitution) supplemented by proliferation. Proliferation and migration may be studied independently by thymidine uptake and proliferation-blocked cell migration using human Caco-2 enterocyte monolayers in culture. Since epidermal growth factor (EGF) promotes mucosal healing and the EGF receptor is a tyrosine kinase, we hypothesized that tyrosine kinases might therefore modulate enterocyte migration and proliferation. The tyrosine kinase inhibitors genistein and 2,5-dihydroxymethylcinnamate, which block kinase ATP-binding and substrate-binding sites, respectively, were studied alone and with EGF. Proliferation was blocked with mitomycin. Although each inhibitor decreased basal and EGF-stimulated monolayer expansion when cell proliferation occurred, neither genistein nor 2,5-dihydroxymethylcinnamate decreased migration when proliferation was blocked. However, each inhibitor prevented EGF stimulation of proliferation-blocked migration and thymidine uptake. More substantial inhibition of basal proliferation by genistein correlated with increased protein-linked DNA breaks, which may reflect nonspecific inhibition of DNA topoisomerase activity by genistein. The more specific 2,5-dihydroxymethylcinnamate blocked changes in the alpha 2 integrin subunit organization which may modulate EGF-stimulated migration. Antiproliferative effects of tyrosine kinase inhibitors decrease basal monolayer expansion but true basal enterocyte migration appears independent of tyrosine kinase regulation. However, a specific tyrosine kinase-dependent modulation of cell-matrix interaction inhibits EGF-stimulated migration.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Division / drug effects
  • Cell Line
  • Cell Movement / drug effects
  • Cinnamates / pharmacology*
  • Dose-Response Relationship, Drug
  • Epidermal Growth Factor / pharmacology*
  • Genistein
  • Humans
  • Immunohistochemistry
  • Intestines / cytology*
  • Intestines / drug effects*
  • Intestines / enzymology
  • Isoflavones / pharmacology*
  • Protein-Tyrosine Kinases / antagonists & inhibitors*

Substances

  • Cinnamates
  • Isoflavones
  • Epidermal Growth Factor
  • methyl 2,5-dihydroxycinnamate
  • Genistein
  • Protein-Tyrosine Kinases