Blockade NF-kappaB activation prohibits TNF-alpha-induced cyclooxygenase-2 gene expression in ED27 trophoblast-like cells.
Among the many functions of trophoblast cells is the production of prostaglandins (PGs) for governing several fetoplacental vascular functions during gestation and the triggering of events leading to parturition. Recent evidence suggests that pro-inflammatory cytokines such as tumour necrosis factors (TNF-alpha) induce PG formation via cyclooxygenase-2 (COX-2), a highly inducible enzyme whose gene is regulated at least in part by inducible transcription factor NF-kappaB. To examine the mechanism by which COX-2-driven PG biosynthesis occurs in trophoblast cells, we utilized the immortalized trophoblast-like cell line ED(27). These cells exhibit many of the properties of villous or extravillous trophoblasts and produce large amounts of PGs in response to TNF-alpha. We demonstrated that challenge of ED(27)cells with TNF-alpha caused binding of the NF-kappaB complex to its kappaB site followed by increased accumulation of COX-2 transcripts. In addition, the inhibitor of NF-kappaB, IkappaB-alpha, became phosphorylated and was rapidly degraded in cytokine-treated cells; this process was abolished by co-incubation with the proteasome inhibitor, MG-132. Finally, when cells were pre-incubated with MG-132 and then challenged with TNF-alpha, PG formation was attenuated in a concentration-dependent manner. These data indicate that, in ED(27)trophoblast-like cells isolated from the first-trimester placenta, TNF-alpha treatment leads to activation of NF-kappaB and subsequent transcription of the COX-2 gene.
Kniss
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DA
Department of Obstetrics and Gynecology (Division of Maternal-Fetal Medicine and Laboratory of Perinatal Research), The Ohio State University, College of Medicine and Public Health, 1654 Upham Drive, Columbus, OH 43210, USA. kniss.1@osu.edu
Rovin
B
B
Fertel
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RH
Zimmerman
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PD
D000328:Adult; D002460:Cell Line; D051546:Cyclooxygenase 2; D004268:DNA-Binding Proteins / Q000378:metabolism; D015232:Dinoprostone / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005260:Female; D006801:Humans; D020672:I-kappa B Proteins; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics; D007976:Leupeptins / Q000494:pharmacology; D008565:Membrane Proteins; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins; D014327:Trophoblasts / Q000201:enzymology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D004268:DNA-Binding Proteins; D020672:I-kappa B Proteins; D007527:Isoenzymes; D007976:Leupeptins; D008565:Membrane Proteins; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D000072000:NF-KappaB Inhibitor alpha; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases; D015232:Dinoprostone; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde