Journal of the Society for Gynecologic Investigation

 

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Journal of the Society for Gynecologic Investigation, Vol. 9, No. 5, 282-289 (2002)
DOI: 10.1177/107155760200900505

Lactosylceramide-Induced Apoptosis in Primary Amnion Cells and Amnion-Derived WISH Cells

Robert M. Moore, MS

David W. Lundgren, PhD

Rebecca J. Silver, BS

Department of Pediatrics, Reproductive Biology, and Biochemistry, Case Western Reserve University, Cleveland, Ohio

John J. Moore, MD

Department of Pediatrics, Reproductive Biology, and Biochemistry, Case Western Reserve University, Cleveland, Ohio; Division of Neonatology, MetroHealth Medical Center, 2500 MetroHealth Drive, Cleveland, OH 441009 jmoore{at}metrohealth.org

Objective: Amnion apoptosis is part of a programmed process of fetal membrane remodeling leading to weakening and rupture. The apoptotis agent lactosylceramide is elevated in amniotic fluid of premature infants with rupture of membranes. We have shown that apoptosis in WISH cells, induced by staurosporine, cycloheximide, or actionomycin D, can be blocked by cyclooxygenase inhibitors, suggesting a relationship between prostaglandin production and apoptosis. Cyclic adenosine monophosphate (cAMP) is known to inhibit prostaglandin release in amnion and WASH cells. This study was undertaken to determine the apoptotic potential of lactosylceramide and the effect of cyclooxygenase inhibitors and cAMP activators on lactosylceramida-induced apoptosis in primary amnion and WISH cells.

Methods: Primary amnion cells and WISH cells were incubated with lactosylceramide to determine apoptosis and prostaglandin E2 (PGE2) release. Apoptosis was confirmed b agarose gel electrophoretic DNA fragmentation analysis, nuclear matrix protein (NMP), and uncleosome enzyme-linked immunosorbent assay. In some studied, cells were preincubated with cyclooxygenase inhibitors or cAMP activators.

Results: Lactosylceramide induced a 20-fold increase in NMP (measure of cell death) in both cell types. Apoptosis was confirmed by the studied listed in methods. Lactosylceramide increased PGE2 release in parallel with apoptosis. Cyclooxygenase inhibitors as well as cAMP activators inhibited both PGE2 release and apoptosis.

Conclusions: Lactosylceramide-induced apoptosis in both amnion and WISH cells. Parallel PGE2 release was demonstrated with apoptosis. Cyclooxygenase inhibitors and cAMP activators blocked both processes.

Key Words: Apoptosis • amnion • lactosylceramide • prostaglandins


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