This Article Full Text (PDF) References Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Wieser, F. Articles by Taylor, R. N. Search for Related Content PubMed PubMed Citation Articles by Wieser, F. Articles by Vigne, J.-L. Articles by Wenzl, R. Articles by Huber, J. Articles by Taylor, R. N. Pubmed/NCBI databases Compound via MeSH Substance via MeSH Social Bookmarking                         What's this?

Effects of Phorbol Dibutyrate on Cell Proliferation, Apoptosis, and Tumor Necrosis Factor-alpha Expression in Human Endometrial Adenocarcinoma Cells

Jean-Louis Vigne, PhD

Rene Wenzl, MD

Johannes Huber, MD, PhD

Division of Gynecological Endocninology and Reproductive Medicine, University of Vienna, Vienna, Austna; Department of Obstetncs and Gynecology, Center for Reproductive Sciences, University of California San Francisco, San Francisco, Califonia.

Robert N. Taylor, MD, PhD

Division of Gynecological Endocninology and Reproductive Medicine, University of Vienna, Vienna, Austna; Department of Obstetncs and Gynecology, Center for Reproductive Sciences, University of California San Francisco, San Francisco, Califonia taylorr{at}obgyn.ucsf.edu

Objectives: Recent evidence suggested that protein kinase C (PKC), a major cell cycle regulator in endometrial models, mimics progesterone withdrawal by inducing downstream signals. In the current study we examined the hypothesis that the PKC activator phorbol 12,13 dibutyrate (PDB) would inhibit cell proliferation and induce apoptosis in two endometrial adenocarcinoma cell (EAC) lines, HEC-1B and Ishikawa cells. We further examined whether the induction of tumor necrosis factor-alpha (TNF-alpha) might mediate these effects.

Methods: EAC lines were cultured under standard and serum-free conditions to study the effects of PDB on cell kinetics. Cell proltferation was determined by cell count using a hemacytometer and by incorporation of ³H thymidine into 100% trichloracetic acid-precipitable DNA. Apoptosis was determined by measuring cytoplasmic histone-associated DNA fragments. Conditioned media concentrations of TNF-alpha were measured by a commercially available enzyme-linked immunosorbent assay (ELISA). EACs were transfected with a-125-bp TNF-alpha promoter luciferase construct and treated with PDB to evaluate transcriptional activation.

Results: Activation of the PKC system with PDB (10 nM) decreased cell proliferation and mitogenesis in EACs. PDB induced apoptosis in both EAC lines. EACs exhibit basal TNF-alpha gene expression and protein secretion and these were increased potently by PDB. However, neutralization of TNF-alpha by addition of anti-TNF-alpha antibodies did not prevent the suppression of mitogenesis, induction of apoptosis, or activation of TNF-alpha gene expression by PDB.

Conclusion: Activation of the PKC system leads to inhibition of cell proliferation, induction of apoptosis, and TNF-alpha expression in EACs. However, apoptosis in this setting does not appear to require TNF-alpha action. EACs provide an informative model to investigate aspects of endometrial epithelial remodeling that may occur under physiologic conditions of progesterone withdrawal.

Key Words: Phorbol dibutyrate • tumor necrosis factor-alpha • apoptosis • human endometrial adenocarcinoma cells • Ishikawa cells • HEC-1B

Journal of the Society for Gynecologic Investigation, Vol. 12, No. 5, 370-375 (2005)
DOI: 10.1016/j.jsgi.2005.02.004


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?