Intestinal trefoil factor controls the expression of the adenomatous polyposis coli–catenin and the E-cadherin–catenin complexes in human colon carcinoma cells

Intestinal trefoil factor controls the expression of the adenomatous polyposis coli–catenin and the E-cadherin–catenin complexes in human colon carcinoma cells

^*Division of Investigative Science, Imperial College of Science, Technology and Medicine, Hammersmith Campus, Du Cane Road, London W12 ONN, United Kingdom; ^†Cancer and Immunogenetics Laboratory, Imperial Cancer Research Fund, Institute of Molecular Medicine, John Radcliffe Hospital, Oxford OX3 9DS, United Kingdom; ^§Imperial Cancer Research Fund, Lincoln’s Inn Fields, London WC2A 3PX, United Kingdom; ^¶Gastrointestinal Cancer Program CC-2218 MCN, Vanderbilt University Medical Center, Nashville, TN 37232-2583; ^‖Department of Pathology, Shiga University of Medical Science, Shiga 520-21, Japan; and ^**Histopathology Unit, Imperial Cancer Research Fund, Lincoln’s Inn Fields, London WC2A 3PX, United Kingdom

Contributed by W. F. Bodmer

Abstract

Intestinal trefoil factor 3 (TFF3) is a member of the trefoil family of peptides, small molecules constitutively expressed in epithelial tissues, including the gastrointestinal tract. TFF3 has been shown to promote migration of intestinal epithelial cells in vitro and to enhance mucosal healing and epithelial restitution in vivo. In this study, we evaluated the effect of recombinant TFF3 (rTFF3) stimulation on the expression and cellular localization of the epithelial (E)-cadherin–catenin complex, a prime mediator of Ca²⁺ dependent cell–cell adhesion, and the adenomatous polyposis coli (APC)–catenin complex in HT29, HCT116, and SW480 colorectal carcinoma cell lines. Stimulation by rTFF3 (10⁻⁹ M and 10⁻⁸ M) for 20–24 hr led to cell detachment and to a reduction in intercellular adhesion in HT29 and HCT116 cells. In both cell lines, E-cadherin expression was down-regulated. The expression of APC, α-catenin and β-catenin also was decreased in HT29 cells, with a translocation of APC into the nucleus. No change in either cell adhesion or in the expression of E-cadherin, the catenins, and APC was detected in SW480 cells. In addition, TFF3 induced DNA fragmentation and morphological changes characteristic of apoptosis in HT29. Tyrphostin, a competitive inhibitor of protein tyrosine kinases, inhibited the effects of TFF3. Our results indicate that by perturbing the complexes between E-cadherin, β-catenin, and associated proteins, TFF3 may modulate epithelial cell adhesion, migration, and survival.

Footnotes

↵ ‡ J.A.E. and M.N. contributed equally to this study.
↵ ‡‡ To whom reprint requests should be addressed. e-mail: m.pignatelli{at}rpms.ac.uk.
ABBREVIATIONS:

TFF3,

intestinal trefoil factor 3;

rTFF3,

recombinant TFF3;

E,

epithelial;

EGF,

epidermal growth factor;

EGFR,

EGF receptor;

APC,

adenomatous polyposis coli;

MBP,

maltose binding protein;

TBST,

Tris buffered saline Tween 20;

FAP,

familial adenomatous polyposis;

PI,

propidium iodide