Cytokines regulate prostaglandin H synthase-1 transcription in human amnion-derived cells.
We have isolated the prostaglandin H synthase-1 (PGHS-1) promoter from the human amnion cell line WISH by long template PCR. The fragment was 1124 base pairs in length and shared a 96% sequence identity with the sequence in GenBank. The putative transcription start site is located 18 bp upstream of the start codon. The sequence is TATA-less, but contains multiple Sp-1 sites and a GC box at -132. The fragment was subcloned into the promoterless reporter construct pBLCAT3 to produce the promoter reporter construct pPGHS1CAT. pPGHS1CAT expression in amnion-derived AV3 cells was inhibited by the pro-inflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin 1-beta (IL-1beta). PGHS-1 mRNA levels however, were unchanged over a 16-h time course with either treatment. These results suggest that PGHS-1 transcription is regulated in a negative manner by cytokines in human amnion-derived cells.
Eykholt
R L
RL
Department of Pharmacology and Clinical Pharmacology, University of Auckland, School of Medicine, Grafton, New Zealand. r.eykholt@auckland.ac.nz
Hansen
W R
WR
Potter
S
S
Marvin
K W
KW
Mitchell
M D
MD
D000650:Amnion / Q000187:drug effects* / Q000378:metabolism*; D001483:Base Sequence; D002460:Cell Line; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D017931:DNA Primers / Q000235:genetics; D005260:Female; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D007527:Isoenzymes / Q000235:genetics* / Q000502:physiology; D008565:Membrane Proteins; D007752:Obstetric Labor, Premature / Q000209:etiology / Q000503:physiopathology; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics* / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D017931:DNA Primers; D007375:Interleukin-1; D007527:Isoenzymes; D008565:Membrane Proteins; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; C496511:PTGS1 protein, human; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases