Polyamines are necessary for normal expression of the transforming growth factor [beta] gene during cell migration. Wang, Jian-Ying, Mary Jane Viar, Ji Li, Hui-Jun Shi, Shirley A. McCormack, and Leonard R. Johnson. Department of Surgery, University of Maryland Medical School and Baltimore, VA Medical Center, Baltimore, MD 21201, Department of Physiology and Biophysics, University of Tennessee College of Medicine, Memphis, TN 38163
APStracts 3:0210G, 1996.
The current study tests the hypothesis that intracellular polyamines are involved in the regulation of the gene expression of transforming growth factor beta (TGF[beta]) during epithelial cell migration after wounding. Administration of [alpha]-difluoromethylornithine (DFMO), a specific inhibitor of ornithine decarboxylase (ODC, the first rate -limiting enzyme for polyamine synthesis), depleted cellular polyamines putrescine, spermidine, and spermine in IEC-6 cells. DFMO also significantly reduced basal levels of TGF[beta] mRNA in unwounded cells. The gene expression of TGF[beta] was dramatically stimulated after wounding a monolayer of the cells not treated with DFMO. TGF[beta] mRNA levels significantly increased from 4 to 12 h after wounding, peaking at 6 h at a level 8 times the pre-wounding control. Increased levels of TGF[beta] mRNA in IEC-6 cells after wounding were paralleled by increase in TGF[beta] content. Depletion of intracellular polyamines in the DFMO-treated cells significantly inhibited the increased expression of the TGF[beta] gene in response to wounding. Cell migration also significantly decreased in the DFMO -treated cells. In the presence of DFMO, exogenous TGF[beta] restored cell migration to normal. These results indicate that 1) polyamine depletion induced by DFMO is associated with decreases in the expression of the TGF[beta] gene and cell migration in IEC-6 cells and 2) exogenous TGF[beta] reverses the inhibitory effect of polyamine depletion on cell migration. These findings suggest that polyamines are required for epithelial cell migration in association with their ability to regulate TGF[beta] gene expression.

Received 6 March 1996; accepted in final form 2 October 1996.
APS Manuscript Number G85-6.
Article publication pending Am. J. Physiol. (Gastrointest. Liver
Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 5 November 1996