prostaglandin-d2 and Glioma

Inflammatory mechanisms may play an important role in the pathogenesis of cisplatin nephrotoxicity. Agonists of the peroxisome proliferator-activated receptor-gamma (PPARgamma), such as rosiglitazone, have been recently demonstrated to regulate inflammation by modulating the production of inflammatory mediators and adhesion molecules. The purpose of this study was to examine the protective effects of rosiglitazone on cisplatin nephrotoxicity and to explore the mechanism of its renoprotection.. Mice were treated with cisplatin with or without pre-treatment with rosiglitazone. Renal functions, histological findings, aquaporin 2 (AQP2) and adhesion molecule expression, macrophage infiltration and tumour necrosis factor-alpha (TNF-alpha) levels were investigated. The effect of rosiglitazone on nuclear factor (NF)-kappaB activity and on viability was examined using cultured human kidney (HK-2) cells.. Rosiglitazone significantly decreased both the damage to renal function and histological pathology after cisplatin injection. Pre-treatment with rosiglitazone reduced the systemic levels of TNF-alpha and down-regulated adhesion molecule expression in addition to the infiltration of inflammatory cells after cisplatin administration. Rosiglitazone restored the decreased AQP2 expression after cisplatin treatment. Pre-treatment with rosiglitazone blocked the phosphorylation of the p65 subunit of NF-kappaB in cultured HK-2 cells. Rosiglitazone had a protective effect via a PPARgamma-dependent pathway in cisplatin-treated HK-2 cells.. These results showed that pre-treatment with rosiglitazone attenuates cisplatin-induced renal damage through the suppression of TNF-alpha overproduction and NF-kappaB activation.

Topics: Anilides; Animals; Apoptosis; C-Peptide; Cell Line; Chromans; Cisplatin; Drug Evaluation, Preclinical; Glioma; Humans; Hypoglycemic Agents; Inflammation; Insulin; Intercellular Adhesion Molecule-1; Kidney; Kidney Diseases; Kidney Function Tests; Kidney Tubules, Proximal; Macrophages; Male; Mice; Mice, Inbred C57BL; Monocytes; PPAR gamma; Prostaglandin D2; Protein Transport; Rosiglitazone; Thiazolidinediones; Transcription Factor RelA; Troglitazone; Tumor Necrosis Factor-alpha

15-Deoxy-(Delta12,14)-prostaglandin J(2) (15d-PGJ(2)) is a naturally occurring cyclopentenone metabolite of prostaglandin D(2) (PGD(2)) and is known as a specific potent ligand for the peroxisome proliferators activator receptor-gamma (PPARgamma). 15d-PGJ(2) inhibits cell growth and induces apoptosis in a number of different cancer cells. However, the underlying mechanism by which 15d-PGJ(2) induces cell death remains to be defined. The present study was undertaken to determine the effect of 15d-PGJ(2) on cell death in A172 human glioma cells. 15d-PGJ(2) caused reactive oxygen species (ROS) generation. 15d-PGJ(2)-induced ROS production and cell death were prevented by the antioxidant N-acetylcysteine. Activation of mitogen-activated protein kinases (MAPK) was not observed in cells treated with 15d-PGJ(2 )and inhibitors of MAPK subfamilies also were not effective in preventing 15d-PGJ(2)-induced cell death. 15d-PGJ(2) treatment caused mitochondrial dysfunction, as evidenced by depolarization of mitochondrial membrane potential. 15d-PGJ(2) induced caspase activation at 24 h of treatment, but the 15d-PGJ(2)-induced cell death was not prevented by caspase inhibitors. The antiapoptotic protein XIAP levels and release of apoptosis inducing factor (AIF) into the cytosol were not altered by 15d-PGJ(2) treatment. Taken together, these findings indicate that 15d-PGJ(2) triggers cell death through a caspase-independent mechanism and ROS production and disruption of mitochondrial membrane potential play an important role in the 15d-PGJ(2)-induced cell death in A172 human glioma cells.

Topics: Brain Neoplasms; Caspases; Cell Death; Cell Line, Tumor; Glioma; Humans; Mitogen-Activated Protein Kinases; Prostaglandin D2; Reactive Oxygen Species

Malignant astrocytomas are among the most common brain tumours and few therapeutic options exist. It has recently been recognized that the ligand-activated nuclear receptor PPARgamma can regulate cellular proliferation and induce apoptosis in different malignant cells. We report the effect of three structurally different PPARgamma agonists inducing apoptosis in human (U87MG and A172) and rat (C6) glioma cells. The PPARgamma agonists ciglitazone, LY171 833 and prostaglandin-J2, but not the PPARalpha agonist WY14643, inhibited proliferation and induced cell death. PPARgamma agonist-induced cell death was characterized by DNA fragmentation and nuclear condensation, as well as inhibited by the synthetic receptor-antagonist bisphenol A diglycidyl ether (BADGE). In contrast, primary murine astrocytes were not affected by PPARgamma agonist treatment. The apoptotic death in the glioma cell lines treated with PPARgamma agonists was correlated with the transient up-regulation of Bax and Bad protein levels. Furthermore, inhibition of Bax expression by specific antisense oligonucleotides protected glioma cells against PPARgamma-mediated apoptosis, indicating an essential role of Bax in PPARgamma-induced apoptosis. However, PPARgamma agonists not only induced apoptosis but also caused redifferentiation as indicated by outgrowth of long processes and expression of the redifferentiation marker N-cadherin in response to PPARgamma agonists. Taken together, treatment of glioma cells with PPARgamma agonists may hold therapeutic potential for the treatment of gliomas.

Topics: Animals; Apoptosis; bcl-2-Associated X Protein; bcl-Associated Death Protein; Cadherins; Carrier Proteins; Cell Division; Cell Survival; DNA Fragmentation; Drug Evaluation, Preclinical; Glioma; Humans; Nuclear Proteins; Oligonucleotides, Antisense; Prostaglandin D2; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-bcl-2; Pyrimidines; Rats; Receptors, Cytoplasmic and Nuclear; Thiazoles; Thiazolidinediones; Transcription Factors; Tumor Cells, Cultured

Tenascin-C (Tn-C) is an extracellular matrix protein with growth-, invasive-, and angiogenesis-promoting activities. Tn-C is upregulated during wound healing, tumorigenesis, and other pathological conditions. Highly malignant gliomas with poor prognosis exhibit high levels of Tn-C expression. Here we demonstrate that Tn-C RNA expression in glioma C6 cells is inhibited in a dose-dependent manner by retinoic acid (RA) and 1,25-dihydroxyvitamin D3 (1,25-D3). No additive or synergistic effects were found. Inhibition is maximum 24 hr after RA or 1,25-D3 treatment, prior to a delayed cytotoxic effect starting at day 4-5 of treatment, and correlates with a reduction in the synthesis of Tn-C protein. Tn-C expression is also inhibited, but to a lesser extent by prostaglandin D2 (PGD2). Furthermore, both RA and 1,25-D3, but not PGD2 abolish the induction of Tn-C by the tumor promoter 12-O-tetradecanoyl phorbol 13-acetate. The inhibition of Tn-C expression might be relevant for the anti-cancer activity of RA and 1,25-D3.

Topics: Animals; Calcitriol; Carcinogens; Drug Synergism; Glioma; Prostaglandin D2; Rats; Receptors, Calcitriol; Tenascin; Tetradecanoylphorbol Acetate; Tretinoin; Up-Regulation

Cell kinetics of cancers have been described in books, texts and other reports, but the correlation with action mechanism of antineoplastic agents has rarely been mentioned in the literature. The action mechanism of the antineoplastic agents such as interferon, ACNU and cisplatin was analyzed with use of propidium iodide and BrdU double staining by flow cytometer. Interferon showed S phase accumulation, ACNU and cisplatin blocked the stage of G(2)M phase. Flow cytometry was useful for the analysis of cell kinetics.

Topics: Antineoplastic Agents; Brain Neoplasms; Cell Cycle; Cisplatin; Flow Cytometry; Glioma; Humans; Interferons; Nimustine; Prostaglandin D2; Tumor Cells, Cultured

The accumulation of inositol phosphates (IPs) in response to prostaglandins (PGs) was studied in NG108-15 cells preincubated with myo-[3H]inositol. As a positive control, bradykinin caused accumulation of IPs transiently at an early phase (within 1 min) and continuously during a late phase (15-60 min) of incubation in the cells. PGD2 and PGF2 alpha did not significantly cause the accumulation of IPs at an early phase but significantly stimulated inositol bisphosphate (IP2) and inositol monophosphate (IP) formation at late phase of incubation. The maximum stimulation was obtained at greater than 10(-7) M concentrations of these PGs, the levels being three-and twofold for IP2 and IP1, respectively. 9 alpha, 11 beta-PGF2 has a slight effect but PGE2 and the metabolites of PGD2 and PGF2 alpha have no effect up to 10(-6)M. The effects of PGD2 and PGF2 alpha were not additive, but the effect of each PG was additive to that of bradykinin at a late phase of incubation. Inositol 1-monophosphate was mainly identified in the stimulation by 10(-5) M PGD2 and 10(-5) M PGF2 alpha, whereas both inositol 1-monophosphate and inositol 4-monophosphate were produced in the stimulation by 10(5) M bradykinin. Depletion of extracellular Ca2+ diminished the stimulatory effect of PGD2 and PGF2 alpha and late-phase effect of bradykinin, but simple Ca2+ influx into the cells by high K+, ionomycin, or A23187 failed to cause such late-phase effects. These results suggest that PGD2 and PGF2 alpha specifically stimulate hydrolysis of inositol phospholipids.

Topics: Animals; Bradykinin; Calcium; Chromatography, High Pressure Liquid; Dinoprost; Dose-Response Relationship, Drug; Egtazic Acid; Glioma; Hybrid Cells; Inositol Phosphates; Isomerism; Neuroblastoma; Prostaglandin D2; Stimulation, Chemical; Time Factors; Tumor Cells, Cultured

Various prostaglandins (PGs) (10 nM-30 microM) were added to NG108-15 cells in culture, and changes in the levels of intracellular cyclic GMP and Ca2+ were investigated. Exposure of the cells to PGF2 alpha, PGD2, and PGE2 (10 microM) transiently increased the cyclic GMP content 7.5-, 3.9-, and 3.1-fold, respectively. Furthermore, the increased levels of cyclic GMP correlated well with the rise in cytosolic free Ca2+ concentrations induced by the PGs. Other PGs (10 microM), including metabolites and synthetic analogs, which had no effect on intracellular Ca2+, failed to increase the cyclic GMP content in the cells. When extracellular Ca2+ was depleted from the culture medium, the PG-induced increase in cyclic GMP level was almost completely abolished. In addition, treatment of the cells with quin 2 tetraacetoxymethyl ester dose-dependently inhibited the PG-induced cyclic GMP formation. The increase in cyclic GMP content caused by treatment of the cells with a high K+ level (50 mM) was completely blocked by voltage-dependent Ca2+ entry blockers, such as verapamil (10 microM), nifedipine (1 microM), and diltiazem (100 microM); however, the PG (10 microM)-induced increase in cyclic GMP content was not affected by such Ca2+ entry blockers. These findings indicate that PG-induced cyclic GMP formation may require the rise in intracellular Ca2+ level and that the voltage-dependent Ca2+ channels may not be involved in the PG-induced rise in Ca2+ content.

Topics: Aminoquinolines; Animals; Calcium; Cyclic GMP; Diltiazem; Dinoprost; Dinoprostone; Fluorescent Dyes; Glioma; Hybrid Cells; Kinetics; Mice; Neuroblastoma; Nifedipine; Potassium; Prostaglandin D2; Prostaglandins; Prostaglandins D; Prostaglandins E; Prostaglandins F; Rats; Tumor Cells, Cultured; Verapamil

The prostaglandins (PG) are known to have various physiological effects. Some series of prostaglandins such as PG D2 have been reported to inhibit growth of tumor cells. In this study, the growth-inhibitory effects of PG A2, PG D2, PG J2 and 6-keto PGE1 were investigated in nude mice receiving subcutaneous transplant of human brain tumor. One to two milligram of prostaglandins was given intraperitoneally every day for three weeks. Tumor volumes were measured twice weekly and the tumor reduction rates (treated/control) were evaluated. T/C rate treated with PG D2 or PG A2 was 50-60% respectively. The effectiveness of PG J2 or 6-keto PGE1 was inferior to that of PG A2 or PG D2. But in the evaluation of antitumor effects of PG J2, we must consider the fact that the activity of PG J2, is liable to be lost. The effect of PG D2 on proliferation of cultured glioma cells was also studied. At concentrations of 10 micrograms/ml, PG D2 strongly inhibited the proliferation of glioma cells. However, precise mechanism of prostaglandin action is presently unknown. Further studies are required to clarify the mechanism of antitumor effects of prostaglandins.

Topics: Alprostadil; Animals; Brain Neoplasms; Drug Screening Assays, Antitumor; Glioma; Humans; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Prostaglandin D2; Prostaglandins; Prostaglandins A; Tumor Cells, Cultured

Prostaglandin (PG) D2 was examined for its effect on the growth of a mouse brain tumor cell line in vitro and in vivo. In this study, we used 203 glioma which had been originally induced by methylcholanthrene in C57BL mice and proved to be subcutaneously transplantable and also maintainable in a cell culture system in vitro. Marked inhibition of cell growth was observed in a PGD2-treated group in vitro at a concentration of 5 micrograms/ml. In the in vivo experiment, intraperitoneal or intratumoral administration of PGD2 (0.5 mg/kg) every day for four weeks was started after subcutaneous transplantation. In a control group, the same amount of ethanol without PGD2 was administered. Inhibition of tumor growth was seen with intratumoral administration, although no inhibition was seen with intraperitoneal administration. Histological examination revealed no remarkable change after PGD2 administration. However, on the DNA histogram, an increment of the G0G1 phase and a decrement of the G2M phase occurred after intratumoral administration of PGD2. It was suggested that local administration of PGD2 might be effective through the inhibition of DNA synthesis.

Topics: Animals; Brain Neoplasms; Cell Division; Cell Line; Cells, Cultured; DNA, Neoplasm; Glioma; Interphase; Male; Mice; Mice, Inbred C57BL; Neoplasm Transplantation; Prostaglandin D2; Prostaglandins D

The cytotoxic and cytokinetic effects of prostaglandin (PG)D2 on human and rat glioma cells were studied in vitro, and the morphological changes occurring in glioma cells following PGD2 treatment were investigated by light and electron microscopy. The cytotoxic effect was evaluated by both colony formation assay and cell growth inhibition. The cytokinetic effect was analyzed by DNA histogram using a flow cytometer. It was found that PGD2 had a dose-dependent cytotoxic effect on the glioma cells and that rat C6 cells were less sensitive to PGD2 than human KY cells. Within 24h after treatment of KY cells with PGD2, a time-dependent cytotoxic effect was also observed. However, the effect of PGD2 on glioma cells was reversible at lower concentration. Investigation of KY cells treated with 2.5 micrograms/ml of PGD2 revealed that the cells in S and G2 + M phases gradually decreased in number and subsequently almost all cells were accumulated in the G0 + G1 phase 12 h later. However, no chronological change of the DNA histogram was obvious in the cells treated with a relatively high concentration of PGD2 (10 micrograms/ml). On the other hand, the treated cells showed more severe morphological changes at a higher concentration of PGD2. Glioma cells treated with PGD2 became round with cytoplasmic vacuolization and blebbing. Thus, PGD2 appears to be useful as an adjuvant chemotherapeutic agent for malignant glioma.

Topics: Animals; Cell Cycle; Cell Division; Cells, Cultured; DNA, Neoplasm; Glioma; Humans; Prostaglandin D2; Prostaglandins D; Rats; Tumor Stem Cell Assay

Five cultured human glioma cell lines were investigated for their reaction to prostaglandin (PG) D2 and E2. In all cases a suppressive effect on DNA synthesis as assessed by 3H-thymidine incorporation was seen with all test substances as early as six hours after the addition of the compounds in doses of usually 10(-5) M. A dose response curve was generated in four cases and showed an estimated ED 50 of about 5 X 10(-6)M. The effect was most pronounced at 12 hours after which the cultures began to recover except those which had been incubated with PGD2. In those cultures which had been exposed to PGD2 virtually no thymidine incorporation was seen after 24 hours and as long as 72 hours. In another set of experiments, the effect of PGD 2, PGE 2, two synthetic PGD 2 analogues, with a chlorine substitution in position 9 (DACl) or with a fluoride substitution in position 9 (DAF) and a synthetic prostacyclin-analogue (Iloprost) was investigated after single and repeated addition of the compounds. A second administration after 12 hours of incubation did not result in a further decrease in 3H-thymidine incorporation like that observed during that first incubation period. In general the cells recovered after 24 hours total incubation time except those which had received PGD 2 or repeated doses of PGE 2. Only in those cells which had been treated with PGD 2, an almost complete blockade of 3H-thymidine incorporation was seen even after the single administration. Parallel evaluation of the cells by flow cytometry showed effects on cell cycle distribution at different times of the incubation.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Cell Division; Cell Line; Dinoprost; Dinoprostone; Glioma; Humans; Prostaglandin D2; Prostaglandins D; Prostaglandins E; Prostaglandins F; Prostaglandins, Synthetic; Thymidine

Glioma C6 cells were incubated with [14C]arachidonate to label membrane phospholipids. Muscimol, a selective gamma-aminobutyric acid A receptor agonist, but not (-)-baclofen, a selective gamma-aminobutyric acid B receptor agonist, stimulates [14C]arachidonate release from C6 cells as a result of hydrolysis of a small pool of phosphatidylcholine and phosphatidylethanolamine by phospholipase A2. This release is facilitated by diazepam and a number of other benzodiazepines such as flunitrazepam, medazepam and midazolam (but very little by clonazepam), although these benzodiazepines per se are inactive in causing the release. In addition to increasing the release of [14C]arachidonate, diazepam in the presence of muscimol promotes the release of [14C] prostaglandin D2. Bicuculline inhibits the action of muscimol and facilitation by diazepam. "Peripheral" benzodiazepine ligand, RO 5-4864 (4'-chlordiazepam) antagonizes the action of diazepam, whereas "central" ligand, RO 15-1788, is inactive. The release of arachidonate metabolites stimulated by muscimol and diazepam is unaffected by Cl- channel blockers, picrotoxin and pentylenetetrazol. Based on these results we propose that in glioma C6 cells (and presumably in normal glia) peripheral benzodiazepine receptor interacts functionally with gamma-aminobutyric acid A type of receptor, which appears not to be linked to picrotoxin sensitive Cl- channel, and may be linked to phospholipase A2.

Topics: Animals; Anti-Anxiety Agents; Arachidonic Acid; Arachidonic Acids; Cell Line; Chromatography, High Pressure Liquid; Diazepam; Drug Interactions; Enzyme Activation; Glioma; Muscimol; Neuroglia; Oxazoles; Phospholipases; Phospholipases A; Phospholipases A2; Prostaglandin D2; Prostaglandins D; Rats; Receptors, GABA-A

The effect of prostaglandin (PG) D2 on neuronal functions was investigated in neuroblastoma X glioma NG108-15 hybrid cells. PGD2 caused a sustained increase in miniature end-plate potentials (MEPPs) recorded from cultured striated muscle cells which had formed junctions with NG108-15 cells. PGD2 initially hyperpolarized and then depolarized NG108-15 cells. The time course of depolarization fitted well to the facilitative phase of MEPPs. The same action on synaptic transmission and membrane potentials was detected with PGF2 alpha but not with PGE1. PGD2 (10(-4)M) produced a 3-fold increase of adenylate cyclase activity in NG108-15 cell homogenates through its receptors that are distinct from those of PGE1 and PGI2. These results show that PGD2 facilitates MEPP frequency from NG108-15 cells due to depolarization, and suggest that PGD2 may act as a physiological neuromodulator for synaptic transmission in vivo.

Topics: Adenylyl Cyclases; Alprostadil; Animals; Cell Line; Dinoprost; Epoprostenol; Glioma; Hybrid Cells; Kinetics; Membrane Potentials; Mice; Muscles; Neuroblastoma; Prostaglandin D2; Prostaglandins D; Prostaglandins E; Prostaglandins F; Rats; Synapses; Synaptic Transmission