Journal Article

Human epidermal keratinocytes undergo (−)-epigallocatechin-3-gallate-dependent differentiation but not apoptosis

Sivaprakasam Balasubramanian, Michael T. Sturniolo, George R. Dubyak and Richard L. Eckert

in Carcinogenesis

Volume 26, issue 6, pages 1100-1108
Published in print June 2005 | ISSN: 0143-3334
Published online February 2005 | e-ISSN: 1460-2180 | DOI: http://dx.doi.org/10.1093/carcin/bgi048
Human epidermal keratinocytes undergo (−)-epigallocatechin-3-gallate-dependent differentiation but not apoptosis

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Epigallocatechin-3-gallate (EGCG) is an important chemopreventive agent derived from green tea. We recently reported that EGCG treatment enhances keratinocyte differentiation as evidenced by increased human involucrin promoter activity [Balasubramanian,S., Efimova,T. and Eckert,R.L. (2002) J. Biol. Chem., 277, 1828–1836]. In the present paper, we extend these findings and show that EGCG also increases the expression of other differentiation markers—procaspase 14 and type I transglutaminase (TG1). Both TG1 mRNA and protein level, and activity are increased by treatment with EGCG. Increased TG1 activity is evidenced by a direct transglutaminase assay, and by the ability of EGCG to stimulate the covalent incorporation of fluorescein cadaverine substrate into crosslinked intracellular structures. In contrast, type II transglutaminase levels are not altered by EGCG treatment. We also assessed whether EGCG promotes keratinocyte apoptosis. We show that EGCG treatment does not promote the cleavage of procaspase-3, -8, -9 or poly(ADP-ribose) polymerase. Moreover, treatment with the pan-caspase inhibitor, Z-VAD-FMK, does not reverse the EGCG-associated reduction in cell viability. In addition, there is no increase in cells having sub-G1/S DNA content, and no evidence for the release of cytochrome c from the mitochondria. These findings confirm, using several endpoints, that EGCG treatment enhances normal keratinocyte differentiation but does not promote apoptosis.

Keywords: COX 4, cytochrome c oxidase; EGCG, epigallocatechin-3-gallate; FC, fluorescein cadaverine; hINV, human involucrin; KSFM, keratinocyte serum-free medium; PARP, poly(ADP-ribose) polymerase; PBS, phosphate-buffered-saline; TG1, type I transglutaminase; TG2, type II transglutaminase

Journal Article.  6647 words.  Illustrated.

Subjects: Clinical Cytogenetics and Molecular Genetics

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