dinoprost and Choriocarcinoma

The 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) is responsible for the inactivation of glucocorticoids. This is the predominant isozyme in the human placenta, where it is proposed to protect the fetus from high levels of maternal cortisol. In the present study, we examined the effects of eicosanoids on the activity of 11beta-HSD2 in human choriocarcinoma JEG-3 cells, a well-established model for placental trophoblasts. Treatment of JEG-3 cells for 24 h with either prostaglandin (PG) E2 or F2alpha attenuated 11beta-HSD2 activity ( approximately 40%). Paradoxically, indomethacin, an inhibitor of cyclooxygenases, inhibited (approximately 40%) rather than stimulated the activity of this enzyme. This indicated that the arachidonic acid metabolism may be diverted to other pathway(s), the products of which may inhibit 11beta-HSD2 activity. To determine whether the lipoxygenase pathways were involved, the cells were treated with nordihydroguaretic acid (NDGA), a blocker of all three (5-, 12-, and 15-) lipoxygenases. NDGA caused a 3-fold increase in 11beta-HSD2 activity. To further delineate which specific lipoxygenase pathway was involved, the cells were incubated with zileuton, a selective inhibitor of 5-lipoxygenase. This resulted in a similar increase in 11beta-HSD2 activity, suggesting that the products of this pathway (e.g., leukotrienes) may be involved. Given that leukotriene B4 (LTB4) is the most biologically active product of the 5-lipoxygenase pathway, we treated the cells with LTB4, which inhibited 11beta-HSD2 activity in a time- and dose-dependent manner with a maximal effect (60% reduction) at 10 nM for 9 h. Semiquantitative reverse transcription-polymerase chain reaction analysis revealed that 11beta-HSD2 mRNA levels were not altered by the addition of LTB4, PGE2, or PGF2alpha, indicating an effect at the posttranscriptional level. In conclusion, these results demonstrate that prostaglandins and LTB4 are potent inhibitors of 11beta-HSD2 activity in JEG-3 cells, suggesting that placental 11beta-HSD2 activity is modulated by these locally produced eicosanoids. This is the first time that the products of arachidonic acid metabolism have been found to regulate the activity of 11beta-HSD2.

Topics: 11-beta-Hydroxysteroid Dehydrogenases; Choriocarcinoma; Dinoprost; Dinoprostone; Enzyme Inhibitors; Humans; Hydroxysteroid Dehydrogenases; Hydroxyurea; Indomethacin; Isoenzymes; Leukotriene B4; Lipoxygenase Inhibitors; Masoprocol; Placenta; Prostaglandins; Tumor Cells, Cultured

Apoptosis has been described in placental (trophoblast) tissues during both normal and abnormal pregnancies. We have studied the effects of the cyclopentenone prostaglandins (PGs) on trophoblast cell death using JEG3 choriocarcinoma cells. PGJ(2), Delta(12)PGJ(2), and 15-deoxy-Delta(12,14)-PGJ(2) (15dPGJ(2)) (10 microM) significantly reduced mitochondrial activity (MTT assay) over 16 h by 17.4 +/- 4.7%, 28 +/- 9.3%, and 62.5 +/- 2.8%, respectively (mean +/- sem), while PGA(2) and PGD(2) had no effect. The synthetic PPAR-gamma ligand ciglitizone (12.5 microM) had a potency similar to 15dPGJ(2) (69 +/- 3% reduction). Morphological examination of cultures treated with PGJ(2) and its derivatives revealed the presence of numerous cells with dense, pyknotic nuclei, a hallmark of apoptosis. FACS analysis revealed an abundance (approximately 40%) of apoptotic cells after 16-h treatment with 15dPGJ(2) (10 microM). The caspase inhibitor ZVAD-fmk (5 microM) significantly diminished the apoptotic effects of Delta(12)PGJ(2) and 15dPGJ(2). JEG3 cells expressed PPAR-gamma mRNA by Northern analysis. These novel findings imply a role for PPAR-gamma ligands in various processes associated with pregnancy and parturition.

Topics: Amino Acid Chloromethyl Ketones; Apoptosis; Choriocarcinoma; Dinoprost; Female; Humans; Labor, Obstetric; Mitochondria; Nuclear Proteins; Pregnancy; Prostaglandin D2; Prostaglandins A; Receptors, Cytoplasmic and Nuclear; RNA, Messenger; Transcription Factors; Transcription, Genetic; Tumor Cells, Cultured; Uterine Neoplasms

The mechanism of labor initiation in humans has not been completely elucidated. Prostaglandins, estrogens, and corticotropin-releasing factor (CRF) have all been shown to affect uterine myocytes and enhance uterine contractility. There are also indications that these uterine regulators have additional effects on other sites involved in labor and that they may act in concert or, perhaps, by regulating each other. Therefore, we evaluated the CRF promoter for transcriptional regulation by prostaglandins and estrogens. Human placental choriocarcinoma cell lines were transfected with CRF-luciferase reporter genes and treated with prostaglandins. Prostaglandin E2 (PGE2), but not prostaglandin F2alpha (PGF2alpha), stimulated CRF-luciferase expression in choriocarcinoma cell lines via a cAMP-dependent pathway. A combination of transfections and in vitro binding studies tested for potential regulation of CRF by estrogen receptor (ER). ER neither regulated the CRF promoter nor interacted with steroid response half-sites from the CRF promoter. Our results provide a molecular regulatory link between PGE2 and CRF, two compounds that enhance uterine contractile function. Combined with the stimulation of prostaglandin release by CRF, these data support a potentially important "feed-forward" regulatory loop involving CRF and PGE2 in parturition. In contrast, we found no evidence for direct effects of estrogens or PGF2alpha on CRF transcription.

Topics: Animals; Cell Line; Choriocarcinoma; Corticotropin-Releasing Hormone; Dinoprost; Dinoprostone; Estradiol; Female; Gene Expression Regulation; Haplorhini; Humans; Luciferases; Placenta; Pregnancy; Prostaglandins; Recombinant Fusion Proteins; Transcription, Genetic; Transfection; Tumor Cells, Cultured; Uterine Neoplasms

The effects of prostaglandin (PG) F2 alpha in 16 patients with vascular malignant intestinal tumors were analyzed by angiography. It was found that PGF2 alpha reduced tumor vascular flow selectively in all but one patient, a rectal carcinoma case. Among the remaining group, a case of intestinal choriocarcinoma complicated by massive gastrointestinal hemorrhage was successfully controlled with intraarterial infusion of PGF2 alpha into the superior mesenteric artery. Owing to the reduced blood flow in tumors, PGF2 alpha is expected to be used extensively as a vasoconstrictor to control bleeding from tumors of the alimentary tract.

Topics: Adult; Aged; Choriocarcinoma; Colonic Neoplasms; Dinoprost; Female; Humans; Intestinal Neoplasms; Intestine, Small; Male; Mesenteric Arteries; Middle Aged; Pregnancy; Prostaglandins F; Radiography; Renal Artery; Uterine Neoplasms