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Cell Structure and Function Dec 2023Calcium transients drive cells to discharge prostaglandin E (PGE). We visualized PGE-induced protein kinase A (PKA) activation and quantitated PGE secreted from a single...
Calcium transients drive cells to discharge prostaglandin E (PGE). We visualized PGE-induced protein kinase A (PKA) activation and quantitated PGE secreted from a single cell by combining fluorescence microscopy and a simulation model. For this purpose, we first prepared PGE-producer cells that express either an optogenetic or a chemogenetic calcium channel stimulator: OptoSTIM1 or Gq-DREADD, respectively. Second, we prepared reporter cells expressing the Gs-coupled PGE reporter EP2 and the PKA biosensor Booster-PKA, which is based on the principle of Förster resonance energy transfer (FRET). Upon the stimulation-induced triggering of calcium transients, a single producer cell discharges PGE to stimulate PKA in the surrounding reporter cells. Due to the flow of the medium, the PKA-activated area exhibited a comet-like smear when HeLa cells were used. In contrast, radial PKA activation was observed when confluent MDCK cells were used, indicating that PGE diffusion was restricted to the basolateral space. By fitting the radius of the PKA-activated area to a simulation model based on simple diffusion, we estimated that a single HeLa cell secretes 0.25 fmol PGE upon a single calcium transient to activate PKA in more than 1000 neighboring cells. This model also predicts that the PGE discharge rate is comparable to the diffusion rate. Thus, our method quantitatively envisions that a single calcium transient affects more than 1000 neighboring cells via PGE.Key words: prostaglandin E, imaging, intercellular communication, biosensor, quantification.
Topics: Animals; Dogs; Humans; HeLa Cells; Dinoprostone; Madin Darby Canine Kidney Cells; Fluorescence Resonance Energy Transfer
PubMed: 37813623
DOI: 10.1247/csf.23047 -
Biochemistry and Cell Biology =... Dec 2023Insensitivity and resistance to 5-fluorouracil (5FU) remain as major hurdles for effective and durable 5FU-based chemotherapy in colorectal cancer (CRC) patients. In...
Insensitivity and resistance to 5-fluorouracil (5FU) remain as major hurdles for effective and durable 5FU-based chemotherapy in colorectal cancer (CRC) patients. In this study, we identified prostaglandin E synthase (PTGES)/prostaglandin E2 (PGE2) axis as an important regulator for 5FU sensitivity in CRC cells. We found that PTGES expression and PGE2 production are elevated in CRC cells in comparison to normal colorectal epithelial cells. Depletion of PTGES significantly enhanced the inhibitory effect of 5FU on CRC cell viability that was fully reverted by exogenous supplement of PGE2. Inhibition of PTGES enzymatic function, by either inducing loss-of-function mutant or treatment with selective inhibitors, phenocopied the PTGES depletion in terms of 5FU sensitization. Mechanistically, PTGES/PGE2 axis modulates glycolysis in CRC cells, thereby regulating the 5FU sensitivity. Importantly, high PTGES expression is correlated with poor prognosis in 5FU-treated CRC patients. Thus, our study defines PTGES/PGE2 axis as a novel therapeutic target for enhancing the efficacy of 5FU-based chemotherapy in CRC.
Topics: Humans; Fluorouracil; Dinoprostone; Prostaglandin-E Synthases; Colorectal Neoplasms; Cell Line, Tumor; Drug Resistance, Neoplasm
PubMed: 37358009
DOI: 10.1139/bcb-2023-0101 -
Prostaglandins, Leukotrienes, and... Dec 2014Previously, we reported that the antidiabetic drug ciglitazone and its analogs were potent inhibitors of 15-hydroxyprostaglandin dehydrogenase (15-PGDH). In continuing...
Previously, we reported that the antidiabetic drug ciglitazone and its analogs were potent inhibitors of 15-hydroxyprostaglandin dehydrogenase (15-PGDH). In continuing attempts to develop highly potent 15-PGDH inhibitors, a series of thiazolidinedione analogs were synthesized and tested. Compound 17 exhibited IC50 of 45 nM. This compound also significantly increased levels of prostaglandin E2 (PGE2) in A549 cells by approximately eight-fold that in the control. Much experimental data suggests that PGE2 plays a role in the prevention of excessive scarring. However, it has a very short half-life in blood, its oxidization to 15-ketoprostaglandins is catalyzed by 15-PGDH. Therefore, 15-PGDH inhibitors may have utility for the therapeutic management of diseases requiring elevated PGE2 levels. Scratch wounds were analyzed in confluent monolayers of HaCaT cells. Cells exposed to compound 17 showed significantly improved wound healing with respect to a control.
Topics: Adenocarcinoma, Bronchiolo-Alveolar; Animals; Benzylidene Compounds; Cell Line; Cell Line, Tumor; Cicatrix; Dinoprostone; Enzyme Inhibitors; Guinea Pigs; Half-Life; Humans; Hydroxyprostaglandin Dehydrogenases; Male; Skin; Thiazolidinediones; Wound Healing
PubMed: 25458900
DOI: 10.1016/j.plefa.2014.09.011 -
Clinical Therapeutics 1993Five hundred fourteen pregnant women at or near term with medically indicated inductions and unfavorable cervical induction features (Bishop score 0-4) were enrolled in... (Clinical Trial)
Clinical Trial Randomized Controlled Trial
Five hundred fourteen pregnant women at or near term with medically indicated inductions and unfavorable cervical induction features (Bishop score 0-4) were enrolled in an open-label randomized multicenter clinical trial to study the effect of endocervical administration of 0.5 mg of dinoprostone cervical gel as a preinduction cervical ripening agent. Patients in the treatment group (n = 265) received dinoprostone cervical gel 12 hours prior to oxytocin induction; patients in the control group (n = 249) were observed during this period. Thirteen patients in each group were excluded from efficacy evaluations. All patients were included in safety analysis. A mean Bishop score increase of 2.9 points was achieved in the treatment group, as compared with 0.6 point in the control group (P < 0.001). During the observation period, spontaneous labor occurred in a significantly greater percentage of patients in the treatment group (27%) than in the control group (2%). The percentage of patients who achieved labor either during the observation period or during the initial induction attempt was significantly larger in the dinoprostone cervical gel group (71.8%) than in the control group (54.2%). In addition, there was a statistically significant (P < 0.001) difference in median induction-to-vaginal delivery time for the treatment group (10.6 hr) and the control group (13.0 hr). Side effects were reported for 42% and 35% of patients in the treatment and control groups, respectively, with fetal heart rate abnormalities reported for approximately 27% of patients in each group.(ABSTRACT TRUNCATED AT 250 WORDS)
Topics: Administration, Intravaginal; Adult; Cesarean Section; Dinoprostone; Female; Gels; Humans; Labor, Induced; Oxytocin; Pregnancy; Time Factors
PubMed: 8269450
DOI: No ID Found -
Dedifferentiation of Human Cardiac Myofibroblasts Is Independent of Activation of COX-2/PGE Pathway.International Journal of Molecular... Mar 2022The differentiation of cardiac fibroblasts to myofibroblasts is considered to be a critical step in activation and progression of cardiac fibrosis in heart disease....
The differentiation of cardiac fibroblasts to myofibroblasts is considered to be a critical step in activation and progression of cardiac fibrosis in heart disease. TGF-β is one of the key cytokines that promotes transition of fibroblasts to myofibroblasts. Dedifferentiation of formed myofibroblasts or reversal of formed myofibroblasts to fibroblasts remains incompletely understood. Prostaglandin E (PGE) has been shown to dedifferentiate human lung myofibroblasts. The role of activation of the COX-2/PGE pathway in dedifferentiation of cardiac myofibroblasts remains unknown. Here, we show that phorbol 12-myristate 13-acetate (PMA) but not PGE induces dedifferentiation of de novo adult human cardiac myofibroblasts stimulated by TGF-β1 from human cardiac fibroblasts as evidenced by reduced expression of α-smooth muscle actin (α-SMA). PMA remarkably increased endogenous levels of PGE in human cardiac myofibroblasts. Pretreatment of myofibroblasts with NS-398, a selective COX-2 inhibitor, and PF-04418948, a selective PGE receptor type 2 (EP2) antagonist, had no effect on expression of α-SMA nor abolished the dedifferentiation induced by PMA. Our results indicated that endogenous and exogenous PGE has no effects on dedifferentiation of cardiac myofibroblasts. PMA-induced dedifferentiation of cardiac myofibroblast is independent of activation of COX-2 and PGE pathway. The mechanism in PMA-induced reversal of cardiac myofibroblasts needs to be explored further.
Topics: Adult; Cell Differentiation; Cells, Cultured; Cyclooxygenase 2; Dinoprostone; Fibroblasts; Heart; Humans; Myofibroblasts; Transforming Growth Factor beta1
PubMed: 35328443
DOI: 10.3390/ijms23063023 -
Journal of Clinical Gastroenterology Dec 1995
Topics: Catheterization; Dinoprostone; Esophagitis, Peptic; Esophagus; Humans
PubMed: 8583096
DOI: 10.1097/00004836-199512000-00002 -
The Journal of Reproductive Medicine Jan 1993The use of locally applied prostaglandin E2 (PGE2) has become a common intervention in the management of the unripened cervix in term pregnancy. The extemporaneously... (Review)
Review
The use of locally applied prostaglandin E2 (PGE2) has become a common intervention in the management of the unripened cervix in term pregnancy. The extemporaneously prepared, vaginally administered prostaglandin E2 gels are most often used clinically in the United States. Large-scale clinical trials have not been conducted with these products. The extemporaneously prepared gels are not standardized with regard to potency, purity or stability; the assessment of efficacy and safety is based on anecdotal clinical reports. PGE2 and its metabolites are eliminated rapidly from the circulation, thus plasma levels are not useful in assessing clinical efficacy. Pharmacokinetic evaluations are based on plasma levels of the stable bicyclo-metabolite of PGE2. Plasma levels resulting from exogenous administration of PGE2 cannot be distinguished from plasma levels caused by endogenous production. Available evidence supports local application of low doses of PGE2 for cervical ripening prior to labor induction. Local application may minimize undesirable systemic effects and uterine hypertonus. A cost/benefit evaluation of a commercially prepared intracervical PGE2 gel must consider several factors in addition to acquisition cost including avoidance of other more costly procedures, such as cesarean section, instrumental delivery, anesthesia, transfusions and monitoring of prolonged labor and fetal outcome, liability issues and pharmacy preparation costs.
Topics: Administration, Intravaginal; Cervix Uteri; Clinical Trials as Topic; Cost-Benefit Analysis; Dinoprostone; Drug Costs; Female; Humans; Infusions, Intravenous; Labor, Induced
PubMed: 8429532
DOI: No ID Found -
Immunologic Research 1989
Review
Topics: Animals; Colony-Forming Units Assay; Dinoprostone; Hematopoiesis; Hematopoietic Stem Cells; Humans; Interleukin-1; Mice
PubMed: 2664032
DOI: 10.1007/BF02918143 -
Experimental Cell Research Dec 2022Prostaglandin metabolism is involved in the regulation of the periodic process of hair follicles. Preliminary research data reported that prostaglandin E2 (PGE2)...
Prostaglandin metabolism is involved in the regulation of the periodic process of hair follicles. Preliminary research data reported that prostaglandin E2 (PGE2) exhibits potential in hair growth. However, the relevant evidence is still insufficient. Herein, we prepared a PGE2 matrix by conjugating PGE2 with collagen via crosslinkers to avoid rapid degradation of PGE2 molecules in vivo. First, we measured the physical properties of the PGE2 matrix. A mouse model of hair loss was established, and PGE2 matrix subcutaneous injection was applied to evaluate hair growth. Under different treatments with the PGE2 matrix, the morphology of hair follicles, the dynamic expression of hair follicle stem cell markers and key regulators in the hair growth cycle were explored. Our data revealed that the PGE2 matrix increased the proportion of developing hair follicles at the early growth stage. Improvements in hair follicle stem cells, such as Sox9+ and Lgr5+ cells, have also been confirmed as therapeutic effects of PGE2 to stimulate hair follicle growth. Our study indicated that PGE2 exhibits effective roles in hair development during anagen. Furthermore, the results also highlight the potential of the PGE2 delivery system as a novel therapeutic strategy for the treatment of hair disorders in the future.
Topics: Mice; Animals; Hair Follicle; Dinoprostone; Hair; Stem Cells; Collagen
PubMed: 36351501
DOI: 10.1016/j.yexcr.2022.113411 -
Journal of Gastroenterology and... Sep 2022Chronic enteropathy associated with the solute carrier organic anion transporter family member 2A1 (SLCO2A1), or CEAS, causes anemia and hypoalbuminemia in young people....
BACKGROUND AND AIM
Chronic enteropathy associated with the solute carrier organic anion transporter family member 2A1 (SLCO2A1), or CEAS, causes anemia and hypoalbuminemia in young people. Dysfunction of the SLCO2A1 transporter protein is thought to involve genetic mutation, but mutant proteins have not been functionally characterized. We examined the prostaglandin E (PGE ) transport ability of recombinant SLCO2A1 proteins containing 11 SLCO2A1 mutations found in CEAS patients.
METHODS
Wild-type and mutant SLCO2A1 proteins were forcibly expressed in Xenopus laevis oocytes, and measurements of PGE uptake and transport capacity were compared. The membrane protein topology and functionality of the eight SLCO2A1 mutations involving single-nucleotide substitutions were predicted using computer analysis.
RESULTS
The extent of functional disruption of the 11 SLCO2A1 mutations identified in CEAS patients was variable, with 10 mutations (421GT, 547GA, 664GA, 770GA, 830dupT, 830delT, 940 + 1GA, 1372GT, 1647GT, and 1807CT) resulting in loss or reduction of PGE transport, excluding 97GC.
CONCLUSION
PGE transport ability of recombinant SLCO2A1 in X. laevis oocytes was hindered in 10/11 SLCO2A1 mutations identified in patients with CEAS. Further studies on the relationships between the different mutations and PGE transport and clinical features, such as severity, are needed.
Topics: Dinoprostone; Humans; Inflammatory Bowel Diseases; Mutation; Organic Anion Transporters
PubMed: 35877192
DOI: 10.1111/jgh.15968