Intestinal trefoil factor induces inactivation of extracellular signal-regulated protein kinase in intestinal epithelial cells

Intestinal trefoil factor induces inactivation of extracellular signal-regulated protein kinase in intestinal epithelial cells

^*Gastrointestinal Unit and Center for the Study of Inflammatory Bowel Disease, Department of Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114; and ^†Biogen, Cambridge, MA 02142

Edited by Kurt J. Isselbacher, Massachusetts General Hospital, Charlestown, MA, and approved November 12, 1997 (received for review July 8, 1997)

Abstract

Intestinal trefoil factor (ITF), a small, compact protease-resistant peptide, is abundantly expressed in goblet cells of large and small intestine. Although several biological activities of ITF have been identified, including promotion of wound healing, stimulation of epithelial cell migration, and protection of intestinal epithelial barrier, little is known about signaling events through which ITF mediates its physiological function. In this study, the effects of exogenous ITF on mitogen-activated protein kinase (MAPK) signaling cascades were examined in IEC-6 cells, a nontransformed intestinal epithelial cell line that does not express endogenous trefoil peptides. Stimulation with ITF resulted in rapid decrease in extracellular signal-related protein kinase (ERK) activity and concomitant reduced ERK tyrosine phosphorylation. ITF also decreased activation of ERK activity induced by either transforming growth factor-α, which links extracellular stimuli to the Ras/Raf/MEK/ERK pathway via the epidermal growth factor receptor, or phorbol 12-myristate 13-acetate, which activates Raf through protein kinase C. ITF-induced inhibition of ERK activity was blocked by an inhibitor of tyrosine and dual-specific phosphatases, sodium orthovanadate. In summary, ITF leads to inhibition of ERK and the MAPK pathway through activation of tyrosine or dual-specific phosphatase.

Footnotes

↵ ‡ To whom reprint requests should be addressed at: Gastrointestinal Unit, Massachusetts General Hospital, Fruit Street, Boston, MA 02114. e-mail: podolsky.daniel{at}mgh.harvard.edu.
This paper was submitted directly (Track II) to the Proceedings Office.
Abbreviations: ERK, extracellular signal-regulated kinase; FCS, fetal calf serum; ITF, intestinal trefoil factor; JNK1, c-Jun NH₂-terminal kinase; MAPK, mitogen-activated protein kinase; MBP, myelin basic protein; MEK, MAP kinase kinase; TGFα, transforming growth factor-α; PMA, phorbol 12-myristate 13-acetate; EGF, epidermal growth factor.