prostaglandin-d2 and Ovarian-Neoplasms

To identify biomarkers that could predict response or lack of response to conventional chemotherapy at the time of diagnosis of high‑grade serous ovarian carcinoma (HGSOC), the present study compared large‑scale gene expression from patients with short or long disease‑free survival times, according to the last cycle of chemotherapy, and validated these findings using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and conventional immunohistochemical (IHC) analysis. Samples were selected for microarray evaluation, at the time of diagnosis, using the following criteria: Identical debulking primary surgery, International Federation of Gynaecology and Obstetrics staging, histological subtype and grade. These were divided into 2 groups, regarding the outcome after 2 years of follow-up. Prostaglandin D2 synthase 21 kDa (brain) (PTGDS) was found to be expressed at a significantly higher level in the tumours of patients with a short disease‑free survival time, and this was validated by RT‑qPCR in all samples. Furthermore, the study evaluated PGD2, the protein product of the PTGDS gene, in a large cohort of 114 HGSOC patients using the Ventana Benchmark automated platform, and IHC positivity was correlated with clinicopathological data and outcome. The global gene expression analysis identified 1,149 genes that were differentially expressed in microarray data, according to the patient outcome. Further analysis RT‑qPCR validated PTGDS gene expression in the same samples (r=0.945; P<0.001). IHC analysis showed an inverse profile, with positivity for PGD2 strongly associated with an increase in disease‑free survival (P=0.009), the absence of relapse (P=0.039) and sensitivity to platinum‑based therapy (P=0.016). Multiple Cox regression showed that IHC evaluation of PGD2 was also a prognostic marker associated with relapse (hazard ratio, 0.37; P=0.002). Overall, the results showed that IHC evaluation of PGD2 is an independent marker of good prognosis in HGSOC. This finding contributes to our understanding of the mechanism of tumour regulation and to investigations into biomarkers that predict response to chemotherapy.

Topics: Adult; Aged; Biomarkers, Tumor; Cystadenocarcinoma, Serous; Cytoreduction Surgical Procedures; Disease-Free Survival; Female; Follow-Up Studies; Humans; Intramolecular Oxidoreductases; Kaplan-Meier Estimate; Lipocalins; Middle Aged; Neoplasm Grading; Ovarian Neoplasms; Ovariectomy; Ovary; Prognosis; Prostaglandin D2

Clinically relevant sirtuin (SIRT) inhibitors may possess antitumor activities. A previous study indicated that 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) exhibited potent anticancer activity by SIRT1 inhibition. Therefore, the aim of the present study was to investigate whether its derivatives (J11-C1 and J19) exhibited anticancer activity against ovarian cancer SKOV3 cells. Cell viability was determined using an MTT assay. Cell cycle arrest, apoptosis and autophagy were determined using flow cytometry or western blot analysis. J11-Cl and J19 were less cytotoxic to SKOV3 cells compared with 15d-PGJ2. Molecular docking studies supported the interactions of 15d-PGJ2, J11-Cl and J19 with various amino acids in SIRT1 proteins. Similar to 15d-PGJ2, J11-C1 and J19 inhibited SIRT1 enzymatic activity and decreased SIRT1 expression levels in a concentration-dependent manner. J11-C1 induced apoptotic cell death more effectively compared with J19, which was associated with markedly decreased expression of the anti-apoptotic molecule B-cell lymphoma 2 (Bcl-2). Furthermore, the levels of light chain 3-Ⅱ (LC3-II) and beclin-1 were clearly induced in SKOV3 cells treated with J11-Cl. Thus, 15d-PGJ2 and its derivatives exhibited anticancer activity possibly by inducing apoptotic or autophagic cell death pathways. Collectively, the results of the present study suggest that 15d-PGJ2 and its derivatives exerted antitumor activity by selectively modulating the expression of genes associated with cell cycle arrest, apoptosis and autophagy. Notably, J11-C1 is a novel candidate SIRT1 inhibitor with anticancer activity.

Topics: Antineoplastic Agents; Autophagy; Beclin-1; Cell Cycle; Cell Line, Tumor; Cell Survival; Dose-Response Relationship, Drug; Female; Gene Expression Regulation, Neoplastic; Humans; Microtubule-Associated Proteins; Models, Molecular; Molecular Docking Simulation; Ovarian Neoplasms; Prostaglandin D2; Signal Transduction; Sirtuin 1

Ovarian cancer accounts for the highest mortality among all gynecological cancers, mainly due to the fast developing chemoresistance. The death ligand TRAIL induces apoptosis and is able to sensitize tumor cells to cytostatic drugs without affecting physiological tissue. Combined treatment of TRAIL and the antidiabetic acting PPARγ ligands was shown to induce apoptosis synergistically in different ovarian cancer cell lines.. To investigate feasible TRAIL-dependent inhibition of proliferation and induction of apoptosis in chemoresistant ovarian cancer cell lines, the drug- and TRAIL-sensitive HEY cell line was utilized to develop subclones with selective resistance against cisplatin, etoposide, docetaxel, paclitaxel, gemcitabine and pemetrexed, as well as against TRAIL as control cell line. Expression of the key factors of the TRAIL signaling pathway, TRAIL receptors 1-4, caspase-8, FLIP and XIAP, was analyzed before and after TRAIL treatment by immunoblotting.. Cell proliferation experiments showed TRAIL-dependent inhibition that was further increased by combination treatment with the PPARγ ligands. Simultaneous exposure of TRAIL and the PPARγ ligands also resulted in enhanced induction of apoptosis even in partial TRAIL-resistant HEY cell lines. In the parental HEY cell line, additional treatment with the PPARγ ligands led to an increased protein expression of DR5 and a further decline of XIAP expression.. Therefore, the combinational treatment with TRAIL and PPARγ ligands might be a promising experimental therapy because the PPARγ ligands, especially d15-PGJ(2), sensitize drug-resistant ovarian cancer cells to TRAIL-induced apoptosis.

Topics: Apoptosis; CASP8 and FADD-Like Apoptosis Regulating Protein; Caspase 8; Cell Line, Tumor; Chromans; Drug Resistance, Neoplasm; Drug Synergism; Female; Humans; Ovarian Neoplasms; PPAR gamma; Prostaglandin D2; Receptors, TNF-Related Apoptosis-Inducing Ligand; Thiazolidinediones; TNF-Related Apoptosis-Inducing Ligand; Troglitazone

15-Deoxy-delta-12,14-prostaglandin-J(2) (15d-PGJ(2)), an arachidonic metabolite and a natural PPARγ agonist, is known to induce apoptosis in tumor cells. In this study, we investigated new therapeutic potentials of 15d-PGJ(2) by determining its anticancer effects in wild-type and doxorubicin-resistant ovarian carcinoma cells. Despite high expression of resistance-inducing genes like MDR1, Bcl2 and Bcl-xl, 15d-PGJ(2) strongly induced apoptosis in doxorubicin-resistant (A2780/AD) cells similar to the wild-type (A2780). This was found to be related to caspase-3/7- and NF-κB pathways but not to its PPARγ agonistic activity. 15d-PGJ(2) also was able to reduce the doxorubicin resistance of A2780/AD cells at low doses as confirmed by the inhibition of gene expression of MDR1 (p-glycoprotein) and SIRT1 (a drug senescence gene). We also investigated effects of 15d-PGJ(2) on cell migration and transformation using a wound-healing assay and morphological analyses, respectively. We found that 15d-PGJ(2) inhibited migration most likely due to NF-κB inhibition and induced transformation of the round-shape A2780/AD cells into elongated epithelial cells due to HDAC1 inhibition. Using a 15d-PGJ(2) analog, we found the mechanism of action of these new activities of 15d-PGJ(2) on SIRT1 and HDAC1 gene expressions and enzyme activities. In conclusion, the present study demonstrates that 15d-PGJ(2) has a high therapeutic potential to kill drug-resistant tumor cells and, the newly described inhibitory effects of this cyclo-oxygenase product on SIRT1 and HDAC will provide new opportunities for cancer therapeutics.

Topics: Antineoplastic Agents; Apoptosis; ATP Binding Cassette Transporter, Subfamily B, Member 1; bcl-X Protein; Cell Line, Tumor; Doxorubicin; Drug Resistance, Neoplasm; Female; Gene Expression Regulation, Neoplastic; Histone Deacetylase Inhibitors; Histone Deacetylases; Humans; NF-kappa B; Ovarian Neoplasms; Prostaglandin D2; Prostaglandin-Endoperoxide Synthases; Proto-Oncogene Proteins c-bcl-2; RNA, Messenger; Signal Transduction; Sirtuin 1; Structure-Activity Relationship; Wound Healing

The antitumor activity of delta 7-prostaglandin A1 (delta 7-PGA1) or delta 12-prostaglandin J2 (delta 12-PGJ2) on human ovarian cancer cell lines resistant to cisplatin (CDDP), doxorubicin (ADR), and L-phenylalanine mustard (l-PAM) was studied in vitro. A2780AD, A2780 (parent cells of A2780AD), 2008DDP, and 2008 cells (parent cells of 2008DDP) were used. The antitumor activities of the drugs were defined with 50% inhibitory concentration (IC50) estimated from growth inhibition curves, which were obtained by an indirect colorimetric method. Drug-resistance ratios obtained from IC50 values, by comparing A2780AD and A2780 cells, were 62.5 for ADR, 4.6 for CDDP, 4.9 for l-PAM, 1.5 for delta 7-PGA1, and 1.8 for delta 12-PGJ2. Those obtained by comparing 2008DDP and 2008 cells were 1.1 for ADR, 16.0 for CDDP, 2.9 for l-PAM, 2.3 for delta 7-PGA1, and 3.2 for delta 12-PGJ2. Thus some human ovarian cancer cells resistant to ADR, CDDP, and l-PAM remain sensitive to antitumor PGs.

Topics: Antineoplastic Agents; Cisplatin; Doxorubicin; Drug Screening Assays, Antitumor; Female; Humans; Melphalan; Ovarian Neoplasms; Prostaglandin D2; Prostaglandins A; Tumor Cells, Cultured

In vitro and in vivo effects of prostaglandin D2 on human ovarian cancer growth were examined by using a cell line, designated HR, derived from ascites of patient with serous cystadenocarcinoma of the ovary. The HR cell proliferation in vitro was dose-dependently inhibited at prostaglandin D2 concentrations between 0.1 and 4.0 micrograms per ml. The results of a 51Cr-release assay seemed to indicate that the inhibition resulted from a direct cytotoxic effect exerted by prostaglandin D2. All DNA, RNA, and protein synthesis by the HR cells was also inhibited in a dose-dependent manner with 48 hr exposure to prostaglandin D2. When nude mice were inoculated with 5 X 10(5) HR cells, the 50% survival time in the untreated group was 52 days. The 50% survival time of nude mice treated with 12 mg (but not 4 mg) of prostaglandin D2 per kg was significantly prolonged to 67 days, in addition to a significant inhibition of the tumor growth. Adjuvant effects of prostaglandin D2 on cisplatin in relation to tumor growth were also studied. Combinations of 0.2 or 0.4 microgram cisplatin per ml and 0.05 or 0.1 microgram prostaglandin D2 per ml, which did not affect the HR cell proliferation alone, resulted in a significant inhibition of cell proliferation. In addition, the tumor take of HR cells by nude mice in groups treated with a combination of cisplatin and prostaglandin D2 was inhibited.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Cisplatin; Cystadenocarcinoma; DNA, Neoplasm; Drug Screening Assays, Antitumor; Female; Humans; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Proteins; Neoplasm Transplantation; Ovarian Neoplasms; Prostaglandin D2; RNA, Neoplasm; Tumor Cells, Cultured

The effects of tumor necrosis factor (TNF) and cyclopentenone prostaglandins (PGA2 and delta 12-PGJ2), singly and in combination, against proliferation in three human gynecologic tumor cell lines (HeLa S3, HHUA, and CAOV-3) were examined in vitro. The HeLa S3 and CAOV-3 cells were unresponsive to TNF, and the HHUA cells exhibited a minimal degree of responsiveness to TNF. The HeLa S3 and HHUA cells were responsive dose-dependently to PGA2, and the CAOV-3 cells were slightly responsive to PGA2. All three cell lines were highly responsive to delta 12-PGJ2. The synergistic antiproliferative effect of TNF and delta 12-PGJ2 appeared in all three cell lines. Pretreatment with delta 12-PGJ2 enhanced the effectiveness of TNF in these cell lines, but not vice versa. A synergistic interaction between TNF and PGA2 was absent in these three cell lines. These results suggest that a combined treatment with TNF and delta 12-PGJ2 could provide a new approach to obtaining increased responses in clinical trials.

Topics: Antineoplastic Combined Chemotherapy Protocols; Cell Division; Drug Synergism; Female; HeLa Cells; Humans; Ovarian Neoplasms; Prostaglandin D2; Prostaglandins A; Prostaglandins, Synthetic; Recombinant Proteins; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha; Uterine Neoplasms

The five stable metabolites [prostaglandin F2 alpha (PGF2 alpha), prostaglandin D2 (PGD2), prostaglandin E2 (PGE2), thromboxane B2 (TXB2), and 6-ketoprostaglandin F1 alpha (6-keto-PGF1 alpha)] of arachidonic acid (AA) via the cyclooxygenase pathway were measured by high-resolution gas chromatography-mass spectrometry in M5076 ovarian reticulosarcoma (M5) homogenates at various times after tumor implantation (Days 15, 18, 21, and 24). Vegetating tumor showed an active AA overall metabolism, which significantly increased during tumor growth. Synthesis of selected products (TXB2, PGD2, and PGE2) increased markedly over time (up to 10.6, 3.5, and 0.9 micrograms/g, respectively). The overall metabolic profile was TXB2 much greater than PGD2 greater than PGF2 alpha greater than 6-keto-PGF1 alpha greater than PGE2 on Day 15 and TXB2 much greater than PGD2 much greater than PGF2 alpha greater than 6-keto-PGF1 alpha on Day 24. TXB2 was also by far the most abundant product of in vitro-cultured M5 cells. Chronic treatment of M5-bearing mice with dazmegrel (UK-38,485), a selective thromboxane synthetase inhibitor (100 mg/kg p.o. daily, from Day 7 to killing), resulted in incomplete TXB2 synthesis inhibition, AA metabolism diversion toward the other prostaglandins, and no effects of tumor growth and metastasis. More frequent dazmegrel treatment (100 mg/kg p.o. every 8 h from Day 1 to killing) resulted in complete TXB2 synthetase inhibition, AA metabolism diversion, and increased tumor growth and metastasis. These data do not support the hypothesis of thromboxane synthetase inhibitors reducing tumor growth. However, since TXB2 suppression was accompanied by the production of other products possibly interfering in tumor growth, no conclusions on the effective role of TXA2 in malignancy can be drawn.

Topics: Animals; Arachidonic Acid; Arachidonic Acids; Dinoprostone; Female; Gas Chromatography-Mass Spectrometry; Imidazoles; Lymphoma, Non-Hodgkin; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Ovarian Neoplasms; Prostaglandin D2; Prostaglandins; Prostaglandins D; Prostaglandins E; Thromboxane B2; Thromboxane-A Synthase; Thromboxanes

Topics: Cell Division; Cell Line; Endorphins; Enkephalin, Methionine; Female; Humans; Melatonin; Naloxone; Ovarian Neoplasms; Prostaglandin D2; Prostaglandins D; Tumor Cells, Cultured

In vitro and in vivo effects of prostaglandin D2 on human ovarian tumor growth were examined by using a cell line, designated HR, derived from ascites of a patient with serous cystadenocarcinoma of the ovary. The HR cell proliferation in vitro was dose dependently inhibited between concentrations of 0.1 and 4.0 micrograms of prostaglandin D2 per ml. From results of 51Cr release assays and trypan blue dye exclusion tests the inhibitory effect seemed to result from a direct cytotoxic effect by prostaglandin D2. All DNA, RNA, and protein syntheses by the HR cells were also inhibited in a dose-dependent manner with exposure time of 48 h to prostaglandin D2. When 5 X 10(5) HR cells were inoculated to nude mice, the 50% survival time of them in untreated groups was 52 days after inoculation. Although 4 mg of prostaglandin D2 per kg caused inhibition of the tumor growth, a significant prolongation of the survival time was not observed. On the other hand, the 50% survival time of nude mice treated with 12 mg of prostaglandin D2 per kg was significantly (P less than 0.05) prolonged to 67 days, in addition to a significant inhibition of the tumor growth.

Topics: Animals; Cell Cycle; Cell Survival; DNA, Neoplasm; Dose-Response Relationship, Drug; Female; Humans; Mice; Mice, Nude; Neoplasm Proteins; Neoplasm Transplantation; Ovarian Neoplasms; Prostaglandin D2; Prostaglandins D; RNA, Neoplasm