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Papers In Press, published online ahead of print November 6, 2001
Physiology and Biophysics, Rm E532, Case Western Reserve Univeristy School of Medicine, Cleveland, Ohio 44106-4970
Corresponding Author: rle2{at}po.cwru.edu
(-)-Epigallocatechin-3-gallate (EGCG) is an important bioactive constituent of green tea that efficiently reduces epidermal cancer cell proliferation. This inhibition is associated with a reduction in activator protein 1 (AP1) transcription factor level and activity. However, its effects on AP1 function in normal epidermal cells have not been extensively explored. Our present studies show that EGCG regulates normal keratinocyte function. To understand the mechanism of action, we examined the effects of EGCG on AP1 factor activity, MAPK signal transduction, and expression of the AP1 factor-regulated human involucrin (hINV) gene. EGCG increases hINV promoter activity in a concentration-dependent manner that requires the presence of an intact hINV promoter AP1 factor binding site. This response appears to be physiologic, as endogenous hINV gene expression is also increased. Fra-1, Fra-2, fosB, junB, junD, c-jun and c-fos levels are increased by EGCG treatment, as is AP1 factor binding to hINV promoter AP1 site. Gel mobility shift studies show that this complex contains Fra-1 and junD. Signal transduction analysis indicates that the EGCG response requires Ras, MEKK1, MEK3 and p38 kinases. Kinase assays and inhibitor studies suggest that p38
J. Biol. Chem, 10.1074/jbc.M110376200
Submitted on October 29, 2001
Revised on November 6, 2001
Accepted on November 6, 2001
Green tea polyphenol stimulates a RAS, MEKK1, MEK3, p38 cascade to increase AP1 factor-dependent involucrin gene expression in normal human keratinocytes
is the p38 isoform responsible for the regulation. These changes are also associated with a cessation of cell proliferation and enhanced cornified envelope formation. These studies show that in normal human keratinocytes EGCG markedly increases, via a MAPK signaling mechanism, AP1 factor-associated responses.
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