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Receptor Expression by Interleukin-1ß in Cultured Human Granulosa-Luteal Cells1
Departments of Obstetrics and Gynecology and Clinical Chemistry (K.N., A.R.) and the Haartman Institute, Department of Bacteriology and Immunology (O.R., A.R), University of Helsinki, Helsinki, Finland
Address all correspondence and requests for reprints to: Dr. Ari Ristimäki, Research Laboratory, Department of Obstetrics and Gynecology, University of Helsinki, Haartmaninkatu 2, SF-00290 Helsinki, Finland
Prostanoids are important regulators of ovarian function, especially
during ovulation and luteolysis. Cyclooxygenase (Cox) is the
rate-limiting enzyme in conversion of arachidonic acid to prostanoids.
We have examined the expression and regulation of the inducible Cox
isoform (Cox-2) and of the receptor for PGF2
(FP) in
human granulosa cells obtained from women undergoing oocyte retrieval
for in vitro fertilization. Freshly isolated granulosa
cells express Cox-2 and FP receptor messenger RNAs (mRNAs). FP receptor
mRNA is also expressed in cultured human granulosa-luteal (GL) cells,
but Cox-2 transcripts are expressed only upon induction.
Interleukin-1ß (IL-1ß) elevated Cox-2 mRNA steady state levels in a
concentration-dependent manner, and kinetic studies showed that Cox-2
mRNA levels were already induced at the 2 h point and returned to
the basal level after incubation for 24 h. The protein synthesis
inhibitor, cycloheximide, induced Cox-2 mRNA expression and potentiated
the effect of IL-1ß. Degradation of Cox-2 mRNA was inhibited by
IL-1ß, which suggests regulation at the posttranscriptional level.
IL-1ß also induced the expression of Cox-2 protein, as detected by
immunofluorescence staining using Cox-2-specific polyclonal antibodies.
Further, IL-1ß-induced synthesis of prostanoids was blocked by a
Cox-2-specific inhibitor, NS-398. In addition, hCG induced Cox-2 mRNA
expression and potentiated the effect of IL-1ß. However, in contrast
to the rapid and transient effect of IL-1ß on Cox-2 mRNA, the effect
of hCG followed slower kinetics. We have previously shown that hCG
induces expression of human FP receptor mRNA in cultured human GL
cells. We now show that IL-1ß induces FP receptor mRNA in a time- and
concentration-dependent manner. Our data suggest that Cox-2 and FP
receptor are coexpressed in freshly isolated human granulosa cells and
that their expression is up-regulated by IL-1ß and hCG in cultured
human GL cells.
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