Lactosylceramide-induced apoptosis in primary amnion cells and amnion-derived WISH cells.
Amnion apoptosis is part of a programmed process of fetal membrane remodeling leading to weakening and rupture. The apoptotic 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 actinomycin 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 WISH cells. This study was undertaken to determine the apoptotic potential of lactosylceramide and the effect of cyclooxygenase inhibitors and cAMP activators on lactosylceramide-induced apoptosis in primary amnion and WISH cells. Primary amnion cells and WISH cells were incubated with lactosylceramide to determine apoptosis and prostaglandin E(2) (PGE(2)) release. Apoptosis was confirmed by agarose gel electrophoretic DNA fragmentation analysis, nuclear matrix protein (NMP), and nucleosome enzyme-linked immunosorbent assay. In some studies, cells were preincubated with cyclooxygenase inhibitors or cAMP activators. Lactosylceramide induced a 20-fold increase in NMP (measure of cell death) in both cell types. Apoptosis was confirmed by the studies listed in methods. Lactosylceramide increased PGE(2) release in parallel with apoptosis. Cyclooxygenase inhibitors as well as cAMP activators inhibited both PGE(2) release and apoptosis. Lactosylceramide-induced apoptosis in both amnion and WISH cells. Parallel PGE(2) release was demonstrated with apoptosis. Cyclooxygenase inhibitors and cAMP activators blocked both processes.
Moore
Robert M
RM
Department ofPediatrics, Cleveland, Ohio 44109, USA. jmoore@metroheath.org
Lundgren
David W
DW
Silver
Rebecca J
RJ
Moore
John J
JJ
D000650:Amnion / Q000166:cytology* / Q000378:metabolism; D015703:Antigens, CD / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D002478:Cells, Cultured; D002518:Ceramides / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D015232:Dinoprostone / Q000378:metabolism; D005260:Female; D006801:Humans; D007790:Lactosylceramides / Q000494:pharmacology*; D034521:Nuclear Matrix-Associated Proteins / Q000378:metabolism; D009707:Nucleosomes / Q000378:metabolism; D011247:Pregnancy; D019311:Staurosporine / Q000494:pharmacology
D015703:Antigens, CD; D002518:Ceramides; D016861:Cyclooxygenase Inhibitors; D007790:Lactosylceramides; D034521:Nuclear Matrix-Associated Proteins; D009707:Nucleosomes; D003609:Dactinomycin; C009744:CDw17 antigen; D003513:Cycloheximide; D000242:Cyclic AMP; D019311:Staurosporine; D015232:Dinoprostone