mocetinostat and Ovarian-Neoplasms

We were the first to report that epithelial ovarian cancer (EOC) cells and tissues express myeloperoxidase (MPO) that is known to play a role in immune surveillance and inflammation by myeloid cells. Additionally, we reported that MPO is colocalized with inducible nitric oxide synthase (iNOS), a key pro-oxidant enzyme, and plays a key role in regulating apoptosis in EOC cells. Whereas myeloid cells express MPO in a dimeric form, intriguingly, here we report the unique expression of only the monomeric form of MPO in EOC cells, tissues, and blood of an ovarian cancer patient. Additionally, we have identified a cell membrane receptor, αV/β1 integrin, that is uniquely expressed by both chemosensitive and chemoresistant EOC cells with significantly higher expression in chemoresistant EOC cells. More importantly, we have demonstrated that monoclonal antibodies against αV/β1 integrin induced cytotoxicity in EOC cells, but not in normal cells, that is also synergistic with conventional chemotherapies. Cytotoxicity of αV/β1 antibodies is due to conformational changes in αV/β1 integrin which prevents monomeric MPO binding to αV/β1 integrin inhibiting the activation of MPO, leading to increased apoptosis. Since normal epithelial cells and macrophages lack monomeric MPO and αV/β1 integrin system, targeting this unique MPO-dependent survival mechanism will selectively eliminate EOC cells and will be the target for developing specific ovarian cancer therapies.

Topics: Carcinoma, Ovarian Epithelial; Epithelial Cells; Female; Humans; Ovarian Neoplasms; Peroxidase; Receptors, Vitronectin

Genital use of talcum powder and its associated risk of ovarian cancer is an important controversial topic. Epithelial ovarian cancer (EOC) cells are known to manifest a persistent prooxidant state. Here we demonstrated that talc induces significant changes in key redox enzymes and enhances the prooxidant state in normal and EOC cells. Using real-time reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay, levels of CA-125, caspase-3, nitrate/nitrite, and selected key redox enzymes, including myeloperoxidase (MPO), inducible nitric oxide synthase (iNOS), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), and glutathione reductase (GSR), were determined. TaqMan genotype analysis utilizing the QuantStudio 12K Flex was used to assess single-nucleotide polymorphisms in genes corresponding to target enzymes. Cell proliferation was determined by MTT proliferation assay. In all talc-treated cells, there was a significant dose-dependent increase in prooxidant iNOS, nitrate/nitrite, and MPO with a concomitant decrease in antioxidants CAT, SOD, GSR, and GPX (

Topics: Apoptosis; Carcinoma, Ovarian Epithelial; Catalase; Cell Line, Tumor; Cell Proliferation; Female; Glutathione Peroxidase; Humans; Mutation; Nitric Oxide Synthase Type II; Ovarian Neoplasms; Oxidative Stress; Peroxidase; Reactive Oxygen Species; Risk Factors; Superoxide Dismutase; Talc

The objective of this study was to determine the expression, and effect of targeting CD11b with a monoclonal antibody in ovarian cancer cells.. CD11b expression was determined in epithelial ovarian cancer (EOC) cell lines and tissues by immunofluorescence and flow cytometry. Cytotoxicity of the CD11b antibody and synergism with chemothearapeutic drugs were determined by the MTT Cell Proliferation Assay in human macrophages, normal ovarian epithelial cells, and in both sensitive and chemoresistant EOC cell lines. Cell migration was assessed with a scratch assay and in vivo effects of the CD11b antibody was assessed with a nude mouse ovarian cancer xenograft model. Data was analyzed with either t-tests or one-way ANOVA.. CD11b was unexpectedly expressed in several EOC lines and tissues, but not normal tissues. Targeting CD11b with its monoclonal antibody resulted in intriguing cytotoxic effects in sensitive and chemoresistant EOC lines, while surprisingly not affecting normal cells. More importantly, the cytotoxicity of the CD11b antibody when combined with chemotherapeutic drugs (cisplatin or docetaxel) was significantly synergistic, in both sensitive and chemoresistant EOC cells. The anti-tumorigenic effect of the CD11b antibody was confirmed in an ovarian cancer nude mouse xenograft model.. Here we identify CD11b as a novel target, which selectively induces cytotoxicity in ovarian cancer cells.

Topics: Animals; Antineoplastic Agents; Antineoplastic Agents, Immunological; Apoptosis; Carcinoma, Ovarian Epithelial; CD11b Antigen; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Cisplatin; Docetaxel; Drug Resistance, Neoplasm; Drug Synergism; Epithelial Cells; Female; Flow Cytometry; Fluorescent Antibody Technique; Humans; In Vitro Techniques; Mice; Mice, Nude; Molecular Targeted Therapy; Neoplasm Transplantation; Neoplasms, Glandular and Epithelial; Ovarian Neoplasms; Ovary; Oxidative Stress; Peroxidase; Taxoids; Xenograft Model Antitumor Assays

Cancer of the ovary is mostly discovered at a late stage and cannot be removed by surgery alone. Therefore surgery is usually followed by adjuvant chemotherapy. However, few reliable biomarkers exist to predict response to chemotherapy of ovarian cancer. Previously, we could demonstrate that IL-17 density is indicative for chemosensitivity. This study focuses on the predictive value of myeloperoxidase (MPO) concerning response to chemotherapy of ovarian cancer.. Biopsies of mostly high-grade primary serous ovarian carcinomas and their matched recurrences were stained with MPO after fixation in formalin and embedding in paraffin. For this staining the technique of tissue-microarray was used. Recurrence within 6 months of the completion of platinum-based chemotherapy was defined as chemoresistance as previously publised. Data for MPO could be analyzed in 92 biopsies.. MPO and IL-17 positive immune cells correlated significantly in biopsies of primary and recurrent carcinomas (r s = 0.41; p = 0.004 and r s = 0.40; p = 0.007, respectively). MPO expression alone did not predict response to chemotherapy, but in multivariate cox regression analysis including age, residual disease, number of chemotherapy cycles, FIGO classification and combined categorized MPO and IL-17 cell densities of primary cancer biopsies, the combination of both immune markers was an independent prognostic factor for recurrence-free survival (p = 0.013, HR = .23, 95CI = 0.07-0.73). There was no chemoresistant patient in the subgroup of MPO + IL-17+, neither in primary nor in recurrent cancer biopsies.. High MPO positive cell density enhances the indicative value of IL-17 for response to chemotherapy in ovarian carcinoma. Although, these results have to be validated in a larger cohort.

Topics: Adult; Aged; Biomarkers, Tumor; Female; Humans; Interleukin-17; Middle Aged; Neoplasm Recurrence, Local; Ovarian Neoplasms; Peroxidase; Platinum; Regression Analysis; Survival Analysis; Tissue Array Analysis; Treatment Outcome

Myeloperoxidase (MPO) is an oxidant generating enzyme normally restricted to myeloid cells, however aberrant MPO expression has been found to occur in non-myeloid cells in some disease states. The functional -463GA promoter polymorphism alters MPO expression levels. The -463G is within an SP1 binding site and is associated with higher gene expression. The G allele is most frequent with ~62% of European populations being GG homozygotes. The GA polymorphism has been associated with risk or survival in a variety of cancers including lung and breast cancer. In this study we determined the frequency of the -463G/A polymorphism in 230 ovarian cancer patients, 75 patients with borderline ovarian tumors, and 299 healthy controls. The GG genotype was found to be overrepresented in patients with early stage ovarian cancer (83.3% GG, p = 0.008) as compared to healthy controls (62% GG), suggesting that MPO oxidants may increase risk. Immunohistochemical analysis revealed MPO expression in a subset of columnar ovarian epithelial carcinoma cells in early stage carcinomas.

Topics: Alleles; Case-Control Studies; Female; Gene Expression; Gene Frequency; Genotype; Humans; Lymph Nodes; Neoplasm Metastasis; Neoplasm Staging; Ovarian Neoplasms; Peroxidase; Polymorphism, Genetic; Promoter Regions, Genetic

The dynamics of the redox-dependent processes in blood plasma, neutrophils and erythrocytes of the patients with ovary cancer of the IIIrd clinical stage by FIGO after polychemotherapy according to the CAP scheme is considered. In the blood plasma and erythrocytes there were estimated the values of protein oxidative modification: carbonyl derivatives at λ = 346 nm, 370 nm, 430 nm and 530 nm; the lipid peroxidation parameters: malonic dialdehyde, dienic conjugates, ketodiens, shiff's bases; the fermentative chain of the antioxidant system: activities of catalase, glutationtransferase and superoxide dismutase. In the peripheral blood neutrophils there were cytochemically determined the myeloperoxidase activity and the number of the active neutrophils in the spontaneous NBTR-test. After the polychemotherapy there were detected higher levels of the protein oxidative modification products and the products of the lipid peroxidation in the blood plasma and erythrocytes of the patients. Simultaneous increase of the activity of the antioxidant enzymes in the blood plasma could be evident of a high level of the lipid antioxidants peroxidation system functioning, whereas the simultaneous decrease of the activity of the antioxidant enzymes in the erythrocytes was indicative of passible development of oxidative stress in them. After the chemotherapy there was observed a significant and reliable decrease of the total number of the neutrophils. After the second course of the chemotherapy the activity of myeloperoxidase in them in the spontaneous NBTR-test as well decreased. Such a dynamics of the redox-depended processes in various components of the blood in the tumour carrier was characteristic of the tumor biological picture and required the use of differential multicomponent antioxidant therapy in patients with ovary cancer.

Topics: Antineoplastic Combined Chemotherapy Protocols; Antioxidants; Catalase; Cisplatin; Cyclophosphamide; Doxorubicin; Erythrocytes; Female; Glutathione Transferase; Humans; Leukocyte Count; Lipid Peroxidation; Malondialdehyde; Neoplasm Staging; Neutrophils; Ovarian Neoplasms; Oxidative Stress; Peroxidase; Schiff Bases; Superoxide Dismutase

Epithelial ovarian cancer (EOC) cells are under intrinsic oxidative stress, which alters metabolic activity and reduces apoptosis. Key oxidative stress enzymes, including myeloperoxidase (MPO) and inducible nitric oxide synthase (iNOS), are upregulated and colocalized in EOC cells. Oxidative stress is also regulated, in part, by superoxide dismutase (SOD) and hypoxia-inducible factor (HIF) 1a. Dichloroacetate (DCA) converts anaerobic to aerobic metabolism and thus was utilized to determine the effects on apoptosis, iNOS, MPO, extracellular SOD (SOD-3), and HIF-1a, in EOC cells. Protein and messenger RNA (mRNA) levels of iNOS, MPO, SOD-3, and HIF-1a were evaluated by immunoprecipitation/Western blot and real-time reverse transcriptase-polymerase chain reaction (RT-PCR), respectively, utilizing SKOV-3 and MDAH-2774 treated with DCA. Apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and caspase 3 assays. Dichloroacetate induced apoptosis, reduced MPO, iNOS, and HIF-1a, whereas increased SOD, in both EOC cell lines. In conclusion, reduction of enhanced oxidative stress-induced apoptosis of EOC cells, which may serve as future therapeutic intervention for ovarian cancer.

Topics: Antineoplastic Agents; Apoptosis; Blotting, Western; Caspase 3; Cell Line, Tumor; Dichloroacetic Acid; Dose-Response Relationship, Drug; Epithelial Cells; Female; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Immunoprecipitation; In Situ Nick-End Labeling; Nitric Oxide Synthase Type II; Ovarian Neoplasms; Oxidation-Reduction; Oxidative Stress; Peroxidase; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Superoxide Dismutase

Resistance to apoptosis is a key feature of cancer cells and is believed to be regulated by nitrosonium ion (NO(+))-induced S-nitrosylation of key enzymes. Nitric oxide (NO), produced by inducible nitric oxide synthase (iNOS), is utilized by MPO to generated NO(+). We sought to investigate the expression of myeloperoxidase (MPO) and iNOS in epithelial ovarian cancer (EOC) and determine their effect on S-nitrosylation of caspase-3 and its activity as well as apoptosis.. MPO and iNOS expression were determined using immunofluorescence in SKOV-3 and MDAH-2774 and EOC tissue sections. S-nitrosylation of caspase-3 and its activity, levels of MPO and iNOS, as well as apoptosis, were evaluated in the EOC cells before and after silencing MPO or iNOS genes with specific siRNA probes utilizing real-time RT-PCR, ELISA, and TUNEL assays.. MPO and iNOS are expressed in EOC cell lines and in over 60% of invasive EOC cases with no expression in normal ovarian epithelium. Indeed, silencing of MPO or iNOS gene expression resulted in decreased S-nitrosylation of caspase-3, increased caspase-3 activity, and increased apoptosis but with a more significant effect when silencing MPO.. MPO and iNOS are colocalized to the same cells in EOC but not in the normal ovarian epithelium. Silencing of either MPO or iNOS significantly induced apoptosis, highlighting their role as a redox switch that regulates apoptosis in EOC. Understanding the mechanisms by which MPO functions as a redox switch in regulating apoptosis in EOC may lead to future diagnostic tools and therapeutic interventions.

Topics: Apoptosis; Caspase 3; Cell Line, Tumor; Epithelial Cells; Female; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Gene Silencing; Humans; Immunohistochemistry; Nitric Oxide Synthase Type II; Ovarian Neoplasms; Peroxidase; RNA, Small Interfering; Transfection; Up-Regulation

To study the clinicopathologic features of granulocytic sarcoma.. The clinical and pathologic findings of 38 cases of granulocytic sarcoma were retrospectively analyzed. Immunohistochemical study was performed and the literature was reviewed.. The age of patients ranged from 2 to 77 years (mean = 43.3 years). The male-to-female ratio was 1.5:1. Major clinical presentations included superficial lymph node enlargement and painful soft tissue mass. Follow-up data were available in 18 patients; and 14 of them died of tumor-related diseases. The average duration of survival of the patients was 16.9 months. Histologically, the tumor cells were relatively uniform in appearance and small to medium in size. The cytoplasm was scanty and pale in color. The nuclei were round or focally irregular, with fine chromatin and inconspicuous nucleoli. Mitosis figures were readily identified. Scattered immature eosinophilic myelocytes were seen. Immunohistochemical study showed that the tumor cells in all cases expressed MPO and CD43. Most cases were also positive for CD68, lysozyme, CD99 and TdT. The staining for CD3, CD20, CD79a, pan-cytokeratin and PLAP were negative.. Granulocytic sarcoma is a known histologic mimicker of non-Hodgkin lymphoma, Ewing sarcoma/PNET and embryonal rhabdomyosarcoma. Detailed morphologic examination, when coupled with immunohistochemical study, is useful in arriving at a correct diagnosis.

Topics: Adolescent; Adult; Aged; Burkitt Lymphoma; Child; Child, Preschool; Diagnosis, Differential; Female; Follow-Up Studies; Humans; Leukosialin; Lymph Nodes; Male; Middle Aged; Muscle Neoplasms; Ovarian Neoplasms; Peroxidase; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Retrospective Studies; Sarcoma, Ewing; Sarcoma, Myeloid; Skin Neoplasms; Survival Rate; Young Adult

The studies determining phagocytic activity, the levels of myeloperoxidase (MPO) and cationic proteins (CP), and the results of the spontaneous and induced NST tests of peripheral neutrophilic granulocytes (NGC) revealed their functional changes in the course of tumor progression in patients with ovarian cancer. There was an increase in the total count of NGC with simultaneous reductions in the levels of CP, the activity of MPO and in the absorptive capacity of NGC at the late clinical stages of the disease. The results of the NST test did not change. The findings suggest that a growing tumor affects NGC reproduction.

Topics: Female; Humans; Neutrophils; Ovarian Neoplasms; Peroxidase; Phagocytosis

This study investigated the immunological factors, such as neutrophils number, the level of myeloperoxidase and IL-12, IL-10, TNF-alpha, IFN-gamma, that additionally might correlate with increased susceptibility to Candida infections in cancer patients. A total of 105 cancer patients were evaluated. Patients were examined twice for Candida colonization and presence of Candida antigen and DNA in bloodstream. Serum concentrations of MPO and selected cytokines were quantified by ELISA. The values for myeloperoxidase were decreased in Candida-colonized as well as deep-infected cancer patients groups, compared to healthy persons. In the group of patients suspected of deep candidiasis, we observed significantly elevated level of IFN-gamma compared to control. In the group of Candida-colonized patients, the concentrations of IL-12, TNF- alpha and IFN-gamma were significantly heightened when compared to control.MPO deficiency seems to be one of the important risk factor for deep candidiasis independently of the neutrophil count. The disturbances in cytokines levels in cancer patients group can be connected with underlying cancer disease, its treatment as well as Candida infection. The decreased level of TNF-alpha, in particular may be connected with Candida invasion.

Topics: Adult; Aged; Candida; Candidiasis; Cytokines; Disease Susceptibility; Female; Humans; Interferon-gamma; Interleukin-10; Interleukin-12; Lung Neoplasms; Male; Middle Aged; Ovarian Neoplasms; Peroxidase; Tumor Necrosis Factor-alpha

One way in which parity and use of oral contraceptives may protect against ovarian cancer is by preventing inflammation and oxidative stress associated with ovulation. Since the genes superoxide dismutase (SOD2), myeloperoxidase (MPO), and NAD(P)H:quinone oxidoreductase 1 (NQO1) are involved in inflammation and oxidative stress, we investigated whether variants of these genes are associated with risk of ovarian cancer.. In a hospital-based case-control study, we compared 125 cases and 193 controls with respect to prevalence of (1) the T-->C (val-->ala) substitution at the -9 position in the signal sequence of SOD2; (2) the G-->A substitution at the -463 position in the promoter region of MPO; and (3) the C-->T (pro-->ser) change in exon 6 of NQO1. Genotyping was done using PCR and gel electrophoresis for MPO and NQO1 and using MALDI-TOF mass spectrometry for SOD2.. For SOD2, women with the TC (val/ala) or CC (ala/ala) genotypes were at increased risk [odds ratio (OR) 2.1, 95% confidence interval (CI) 1.1-4.0]. Results for MPO and NQO1 were in the hypothesized directions but were not statistically significant. For MPO, there was a small inverse association among women with GA or AA genotypes (OR = 0.72, 95% CI 0.43-1.2). For NQO1, the TT (ser/ser) genotype was associated with somewhat increased risk (OR = 2.3, 95% CI 0.69-7.6).. While these results need to be confirmed in other studies, they point to a possible role for genes involved in oxidative stress in the development of ovarian cancer.

Topics: Adult; Alleles; Case-Control Studies; Female; Genetic Variation; Genotype; Humans; Middle Aged; NAD(P)H Dehydrogenase (Quinone); Ovarian Neoplasms; Oxidative Stress; Peroxidase; Polymorphism, Genetic; Risk Factors; Superoxide Dismutase

Oxidative stress has been implicated in several diseases, including cancer. Oxidants induce oncogenes and their products associated with cell growth. Even though epidemiological studies implicate oxidants in promoting cancer, there is still a lack of in vivo evidence for the same. In this study, we measured the levels of myeloperoxidase (MPO), an enzyme associated with oxidation and autoantibodies to lipid peroxide-modified protein (LOOH-RSA), in the plasma of subjects with gynecological cancers. The gynecological cancer subjects (n = 201) had higher plasma MPO and LOOH-RSA levels compared with control subjects (n = 60). Immunohistochemical analysis of tissues revealed that immunostaining for MPO and LOOH-RSA was higher in cancer tissues compared with controls. The staining was specific to cell types and not ubiquitously present. Neutrophils, monocytes/macrophages, and natural killer cells have been proposed to play a role in cancer promotion and progression. This study proposes a role for oxidative stress and especially MPO in cancer.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Endometrial Neoplasms; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunohistochemistry; Middle Aged; Ovarian Neoplasms; Oxidative Stress; Peroxidase; Protein Biosynthesis; Proteins; Uterine Cervical Neoplasms

Immunohistochemistry is used for in situ detection of proteins in histological slides and is now an important diagnostic tool. Due to several methodological and biological factors, conventional immunohistochemical procedures may sometimes have too low sensitivity, especially for tracing the histogenesis of malignant tumours. A few years ago, an amplification technique was introduced which greatly increased the sensitivity of some of the commonly used immunohistochemical methods. This technique permits the use of primary antibodies in significantly lower concentrations compared with the conventional methods. Alternatively, one can keep the antibody concentration unchanged and use the enhanced sensitivity to detect scarce proteins, which are not visualised by traditional immunohistochemical procedures. We present a brief description of the technique and show some examples of its use in the diagnosis of neuroendocrine carcinomas. Tyramide signal amplification might become an important supplement for the diagnosis and classification of malignant tumours.

Topics: Avidin; Biotin; Bronchi; Carcinoma, Small Cell; Endocrine Gland Neoplasms; Female; Humans; Immunohistochemistry; Lung Neoplasms; Ovarian Neoplasms; Peroxidase; Sympathomimetics; Tyramine

Topics: Base Sequence; Breast; Breast Neoplasms; Chromosome Walking; Chromosomes, Artificial, Yeast; Chromosomes, Human, Pair 17; DNA-Binding Proteins; Female; Genes; Humans; Molecular Sequence Data; Multigene Family; Ovarian Neoplasms; Ovary; Peroxidase; Sequence Tagged Sites; Transcription Factors; Tripartite Motif Proteins; Ubiquitin-Protein Ligases