endothelin-1 and Carcinoma

Endothelin axis deregulation triggers a series of events that lead to a profound deregulation in cancer cells, including key tumorigenic cellular events such as proliferation, invasion, escape from programmed cell death, new vessel formation, abnormal osteogenesis and the alteration of nociceptive stimuli. Atrasentan is a novel agent that effectively targets this pathway and is able to inhibit and/or reverse several of those events. Biological and clinical activity in patients with prostate cancer has been demonstrated in a Phase III clinical setting by the suppression of markers of biochemical and clinical prostate cancer progression, and by a delay in time to disease progression, especially in patients with bone disease.

Topics: Atrasentan; Carcinoma; Clinical Trials, Phase III as Topic; Endothelin-1; Humans; Male; Prostatic Neoplasms; Pyrrolidines; Receptor, Endothelin A

The purpose of this paper is to assess the relationship between gene polymorphism in angiogenesis-related genes and radiation responses in nasopharyngeal carcinoma (NPC) patients. The genotypes of 180 NPC patients were analyzed by Sequenom MassARRAY. The response evaluation criteria in solid tumours were used for assessing efficacies, and the criteria of the Radiation Therapy Oncology Group or European Organization for Research & Treatment of Cancer were utilized for evaluating acute toxic reactions in response to radiation. Statistical methods included chi-square test, uni- and multivariate logistic regression analyses. Genotypic carriers of rs1800541 GT were at an elevated risk of developing grade 3+ oral mucositis, and a genetic variant of rs5333 was a predictor for a lower occurring risk of grade 2+ radiation-induced xerostomia. EDN1 rs1800541, rs2071942 and rs5370 variants were associated with a significantly higher risk of severe myelosuppression. SNPs in such angiogenesis-related genes as EDN1 rs1800541, rs2071942 & rs5370 and EDNRA rs5333 may serve as useful biomarkers for predicting the outcomes of NPC patients.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Carcinoma; Endothelin-1; Female; Humans; Male; Middle Aged; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Neovascularization, Pathologic; Polymorphism, Single Nucleotide; Treatment Outcome; Young Adult

Endothelin-1 (ET-1) and its receptors (ETAR and ETBR), referred to as the endothelin (ET) axis, are overexpressed in breast carcinomas and appear to influence tumour growth and progression. The objective of this study was to determine the effect of expression of the ET axis in breast carcinomas on response to cytotoxic chemotherapy. The study included 44 patients with locally advanced breast cancer participating in a prospective phase III study evaluating high-dose neoadjuvant chemotherapy of epirubicin and cyclophosphamide. Expression of ET-1, ETAR and ETBR was determined by semiquantitative immunohistochemical analysis of breast cancer tissue from prechemotherapy tru-cut biopsies. Immunohistochemical staining was positive for ET-1 in 61.5%, for ETAR in 35% and for ETBR in 35.9% of breast carcinomas. Pathological response to chemotherapy was significantly decreased in ETAR-positive patients (P=0.002). In total, 50% of ETAR-positive patients as compared to 7.7% of ETAR-negative patients attained pathologically 'no change'. Logistic regression confirmed ETAR as an independent predictive marker for pathological response (P=0.009). These data indicate that increased expression of ETAR in breast carcinomas is associated with resistance to chemotherapy. Determination of ETAR status may serve as a predictive marker for identifying patients less likely to be responsive to conventional chemotherapy.

Topics: Adult; Aged; Antineoplastic Combined Chemotherapy Protocols; Breast Neoplasms; Carcinoma; Chemotherapy, Adjuvant; Cyclophosphamide; Drug Resistance, Neoplasm; Endothelin-1; Epirubicin; Female; Gene Expression Profiling; Humans; Immunohistochemistry; Middle Aged; Neoadjuvant Therapy; Predictive Value of Tests; Receptor, Endothelin A; Receptor, Endothelin B; Sensitivity and Specificity; Treatment Outcome

Epstein-Barr virus-associated gastric carcinoma (EBVaGC) is one of the four subtypes of gastric carcinoma and its unique clinicopathological mechanism is unclear. Herein, the expression of endothelin-1 (ET-1) in EBVaGC was lower than of Epstein-Barr virus-negative gastric carcinoma (EBVnGC) and associated with a low frequency of lymph node metastasis of EBVaGC. Functional studies showed that the activation of ET-1/endothelin receptor type A (ETAR) axis could promote cell growth, migration, and antiapoptosis. The expression of the ET-1 gene was unrelated to methylation of its promoter region and miRNAs (-1, -125a, -125b). After being treated with MEK1/2 inhibitor (PD0325901), the inactivation of ERK1/2 pathway resulted in downregulation of ET-1 and forkhead box O1 (FOXO1) expression. Further, FOXO1 knockdown decreased the ET-1 expression. These findings indicated that ET-1 could be involved in development of gastric cancer and EBV could suppress the expression of ET-1 via the regulation of the transcription factor FOXO1 through the MAPK/ERK pathway.

Topics: Carcinogenesis; Carcinoma; Endothelin-1; Epstein-Barr Virus Infections; Forkhead Box Protein O1; Herpesvirus 4, Human; Humans; MAP Kinase Signaling System; Stomach Neoplasms

Pathological proliferation of human airway smooth muscle cells (HASMCs) causes hyperplasia in chronic lung diseases. Signaling pathways that link airway inflammation to HASMC proliferation might provide therapeutic targets for the prevention of airway remodeling and chronic lung diseases. Endothelin-1 (ET-1) signals via endothelin-A- and B-receptors (ETAR, ETBR) to perpetuate HASMC-associated and TNFα-dependent inflammatory processes.. endothelin receptor antagonists (ERAs) suppress HASMC proliferation induced by inflammatory cytokines. HASMCs were stimulated ex vivo with cytokines in the presence or absence of ERAs (ETAR-specific/selective: BQ123, ambrisentan; ETBR-specific: BQ788; non-selective: bosentan, macitentan, ACT-132577) or cytokine-blocking antibodies. Cell counts, DNA-synthesis (BrdU-incorporation assay), cytokine production (ELISA) and ETBR expression (whole-genome microarray data, western blot) were analyzed. ET-1-induced HASMC proliferation and DNA-synthesis were reduced by protein kinase inhibitors and ETAR-specific/selective ERAs but not by BQ788. TNFα-induced HASMC proliferation and DNA-synthesis were reduced by all ERAs. TNFα induced ET-1 and ETBR expression. TNFα- and ET-1-induced GM-CSF releases were both reduced by BQ123 and BQ788. TNFα- and ET-1-induced IL-6 releases were both reduced by BQ123 but not by BQ788. Combined but not single blockade of GM-CSF-receptor-α-chain and IL-6 reduced TNFα- and ET-1-induced HASMC proliferation and DNA-synthesis. Combined but not single treatment with GM-CSF and IL-6 induced HASMC proliferation and DNA-synthesis in the presence of ET-1. In conclusion, TNFα induces HASMC proliferation via ET-1/GM-CSF/IL-6. ETBR requires up-regulation by TNFα to mediate ET-1 effects on HASMC proliferation. This signaling cascade links airway inflammation to HASMC-associated remodeling processes and is sensitive to ERAs. Therefore, ERAs could prevent inflammation-induced airway smooth muscle hyperplasia.

Topics: Antibodies, Blocking; Biomarkers; Bronchi; Bronchial Neoplasms; Carcinoma; Cell Proliferation; Cells, Cultured; DNA Replication; Endothelin Receptor Antagonists; Endothelin-1; Gene Expression Regulation; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Hyperplasia; Interleukin-6; Muscle, Smooth; Protein Kinase Inhibitors; Receptor, Endothelin A; Receptor, Endothelin B; Recombinant Proteins; Signal Transduction; Tumor Necrosis Factor-alpha

To explore the role of endothelin-1 (ET-1) gene in regulating the proliferation, migration and invasion of nasopharyngeal carcinoma cells.. A lentivirus-mediated shRNA-ET-1 vector was infected into 5-8F cells, and the interference efficiency was examined with Western blotting. MTT assay, cell cycle analysis, plate colony formation assay, Transwell assay, Boyden chamber assay and tumor growth assay were carried out to analyze the changes in cell proliferation, migration and invasion. The expressions of genes related with epithelial-mesenchymal transition (EMT) were examined using Western blotting.. shRNA-ET-1 transfection significantly inhibited the expression of ET-1, and suppressed the growth, migration and invasion of 5-8F cells. ET-1 knockdown enhanced the expression of E-cadherin and CK18 and inhibited the expression of N-cadherin and vimentin.. ET-1 promotes cell growth, migration and invasion by modulating the genes associated with epithelial-mesenchymal transition in nasopharyngeal carcinoma cells.

Topics: Antigens, CD; Cadherins; Carcinoma; Cell Cycle; Cell Line, Tumor; Cell Movement; Cell Proliferation; Endothelin-1; Epithelial-Mesenchymal Transition; Gene Silencing; Humans; Lentivirus; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Neoplasm Invasiveness; RNA, Small Interfering; Transfection; Vimentin

Endothelins and their receptors have been implicated in numerous diseases and have recently emerged as relevant players in a variety of malignancies. Tumours overexpress the Endothelin-1 (ET-1) and the Endothelin-A receptors (ETAR) and their interaction enhances tumour growth and metastasis by promoting tumour cell survival, proliferation and angiogenesis. In this study we have evaluated the expression of ET-1 and ETAR in 50 canine mammary tumours, compared to normal controls. Results demonstrated a progressive increase in ET-1 and ETAR expression from benign tumour to grade 1 and to grade 2 malignant mammary tumours with a decrease of expression in grade 3 carcinomas. Co-localization of ET-1 and ETAR was observed in benign mammary tumours and in G1 and G2 carcinomas, while absent in G3 carcinomas. Concluding, ET-1/ETAR can be considered reliable markers for evaluating malignancy of canine mammary tumours and could have importance for the development of specific anticancer therapies.

Topics: Animals; Blotting, Western; Carcinoma; Dog Diseases; Dogs; Endothelin-1; Female; Fluorescent Antibody Technique; Immunohistochemistry; Mammary Neoplasms, Animal; Receptor, Endothelin A

In order to become established in the skeleton, metastatic cells disseminating from the breast carcinoma need to acquire organ-specific traits. There are no effective predictors for who will develop bone metastasis to guide long-term predictive therapy. Our purpose was to individuate events critical for bone colonization to make a molecular classification of breast carcinoma useful for bone-metastasis outcome. In dysplasia adjacent to carcinoma and in pair-matched specimens of bone metastasis we examined SPARC expression and localization as well as Endothelin 1/ETAR signals by immunohistochemistry, and the evaluation of plasma levels of SPARC by ELISA was also performed. In patients with breast carcinoma metastasizing to bone, SPARC and Endothelin 1/ETAR axis were highly expressed from dysplasia until bone metastasis, but the SPARC plasma level was as low as that of normal women, in contrast to patients that never develop bone metastasis, suggesting that circulating SPARC was counter adhesive. Altogether, the early identification of SPARC/Endothelin 1/ETAR in dysplastic lesions would be important to devise therapies preventing metastasis engraftment, since often carcinoma cells spread to distant organs at the time or even before patients present with cancer.

Topics: Adult; Biomarkers, Tumor; Bone Neoplasms; Breast Neoplasms; Carcinoma; Case-Control Studies; Endothelin-1; Female; Humans; Middle Aged; Osteonectin; Receptor, Endothelin A

Emerging evidence clearly indicates that EZH2 plays a crucial role in tumor angiogenesis. However, the role of EZH2 in angiogenesis is still unknown in nasopharyngeal carcinoma (NPC). We here showed that the elevated EZH2 level was closely associated with an aggressive and poor prognostic phenotype, and was positively correlated with microvessel density (MVD) in NPC tissues. Functional studies showed that EZH2 upregulation promoted cell proliferation, migration and tubule formation of endothelial cells, and knockdown of EZH2 suppressed tumor growth, metastasis and angiogenesis in vivo. Mechanistic investigations revealed that EZH2 inhibited miR-1 transcription via promoter binding activity, leading to enhanced expression of Endothelin-1 (ET-1) which is suppressed by miR-1 targeting of ET-1 3'UTR. Furthermore, knockdown of EZH2 or overexpression of miR-1 exerted anti-angiogenic effect on NPC cells. More importantly, the neutralizing antibody against ET-1 significantly abrogated the pro-angiogenic effect of EZH2, and forced expression of ET-1 rescued the anti-angiogenic effect induced by EZH2 knockdown. In clinical specimens, ET-1 was widely overexpressed and associated with clinical stage and MVD. Taken together, our results identify a novel signaling pathway involved in NPC angiogenesis, and also suggest that EZH2-miR-1-ET-1 axis represents multiple potential therapeutic targets for NPC.

Topics: 3' Untranslated Regions; Angiogenesis Inhibitors; Animals; Base Sequence; Carcinoma; Cell Line, Tumor; Cell Proliferation; Chick Embryo; Endothelin-1; Enhancer of Zeste Homolog 2 Protein; Gene Knockdown Techniques; Heterografts; Human Umbilical Vein Endothelial Cells; Humans; Mice; MicroRNAs; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Neovascularization, Pathologic; Polycomb Repressive Complex 2; Transfection

Here, we report a complex regulation of endothelin-1 (ET-1) axis driven by epigenetic reactions in 1833-bone metastatic cells, emphasizing the importance in skeletal metastasis from breast carcinoma. Inhibitors of histone deacetylases, trichostatin A (TSA), and of DNA methylases, 5'-Azacytidine (Aza), caused, respectively, reduction and increase in 1833 cell invasiveness, without affecting the basal migration of parental MDA-MB231 cells. Of note, in the two cell lines exposed to Aza the blockade of the ET-1 receptor ETAR with BQ-123 oppositely changed invasive properties. Even if in MDA-MB231 cells the ET-1 axis was scarcely influenced by epigenetic reactions, ETAR remarkably decreased after Aza. In contrast, in 1833 cells Aza exposure enhanced ET-1 coupled to ETAR wild type, being also ETAR truncated form increased, and invasiveness was stimulated. Under demethylation, the increase in ET-1 steady state protein level in 1833 clone seemed regulated at transcriptional level principally via Ets1 transcription factor. In fact, actinomycin D almost completely prevented ET-1 mRNA induction due to Aza. Only in 1833 cells, TSA exposure inactivated ET-1 axis, with reduction of the expression of ET-1 and ETAR mutated form, in agreement with Matrigel invasion decrease. This treatment favoured the ET-1 repressional control, taking place at the level of mRNA stability due to the 3'-untranslated region in the ET-1 gene, and also decreased transcription via NF-kB. Environmental conditions that alter the balance between epigenetic reactions might, therefore, affect metastasis migratory mode influencing ET-1 axis.

Topics: Azacitidine; Bone Neoplasms; Breast Neoplasms; Carcinoma; Cell Line, Tumor; Cell Movement; Collagen; Dactinomycin; DNA Methylation; Drug Combinations; Endothelin A Receptor Antagonists; Endothelin-1; Enzyme Inhibitors; Epigenesis, Genetic; Female; Gene Expression Regulation, Neoplastic; Histone Deacetylase Inhibitors; Humans; Hydroxamic Acids; Laminin; Mutation; Neoplasm Invasiveness; Peptides, Cyclic; Proteoglycans; Proto-Oncogene Protein c-ets-1; Receptor, Endothelin A; RNA Stability; RNA, Messenger; Transcription, Genetic

Recent studies have indicated that the expression of endothelin A receptor (ETAR) and chemokine receptor 4 (CXCR4) could be used as an indicator of the metastatic potential of nasopharyngeal carcinoma (NPC). The aim of this study was to determine the prognostic value of ETAR and CXCR4 in NPC patients and to reveal the interplay of the endothelin-1 (ET-1)/ETAR and stromal-derived factor-1(SDF-1)/CXCR4 pathways in promoting NPC cell motility.. Survival analysis was used to analyze the prognostic value of ETAR and CXCR4 expression in 153 cases of NPC. Chemotaxis assays were used to evaluate alterations in the migration ability of non-metastatic 6-10B and metastatic 5-8F NPC cells. Real-time PCR, immunoblotting, and flow cytometric analyses were used to evaluate changes in the expression levels of CXCR4 mRNA and protein induced by ET-1.. The expression levels of ETAR and CXCR4 were closely related to each other and both correlated with a poor prognosis. A multivariate analysis showed that the expression levels of both ETAR and CXCR4 were independent prognostic factors for overall survival (OS), progression-free survival (PFS), and distant metastasis-free survival (DMFS). The migration of 6-10B and 5-8F cells was elevated by ET-1 in combination with SDF-1α. The knockdown of ETAR protein expression by siRNA reduced CXCR4 protein expression in addition to ETAR protein expression, leading to a decrease in the metastatic potential of the 5-8F cells. ET-1 induced CXCR4 mRNA and protein expression in the 6-10B NPC cells in a time- and concentration-dependent fashion and was inhibited by an ETAR antagonist and PI3K/AKT/mTOR and MAPK/ERK1/2 pathway inhibitors.. ETAR and CXCR4 expression levels are potential prognostic biomarkers in NPC patients. ETAR activation partially promoted NPC cell migration via a mechanism that enhanced functional CXCR4 expression.

Topics: Carcinoma; Cell Differentiation; Cell Line, Tumor; Cell Movement; Chemokine CXCL12; Chemotaxis; Endothelin-1; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Male; MAP Kinase Signaling System; Middle Aged; Multivariate Analysis; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Prognosis; Proto-Oncogene Proteins c-akt; Receptor, Endothelin A; Receptors, CXCR4; RNA, Messenger; RNA, Small Interfering; Up-Regulation

We aimed to investigate the prognostic role of endothelin-1 (EDN1) and endothelin A receptor (EDNRA) gene polymorphisms in patients with locoregionally advanced nasopharyngeal carcinoma (NPC).. Two hundred three consecutive patients with locoregionally advanced NPC were enrolled. Seven potentially functional polymorphisms in the EDN1 and EDNRA genes were determined by ligase detection reaction-PCR method from prospectively collected blood samples. The influence of the genetic polymorphisms on patient overall survival (OS) was analyzed using Cox proportional hazards model, Kaplan-Meier method, and the log-rank test.. The 5-year OS in patients with EDNRA/H323H TT, TC, and CC genotypes were 81.3%, 62.1%, and 75.0%, respectively (P = 0.004). Patients carrying the heterozygous (TC) or homozygous variant (CC) genotype in EDNRA/H323H were combined for analysis, which revealed that the 5-year OS in patients with TC/CC genotypes was significantly lower than those with the wild-type TT genotype (63.2% vs. 81.3%; P = 0.002). Multivariate analysis showed that EDNRA/H323H polymorphism (HR: 1.95; 95% CI: 1.18-3.23; P = 0.009) and N classification (HR: 1.35; 95% CI: 1.03-1.79; P = 0.03) were independent significant prognostic factors for OS in patients with locoregionally advanced NPC. In contrast, the EDN1 polymorphisms revealed no prognostic value.. The EDNRA/H323H polymorphism was a novel and independent prognostic marker for patients with locoregionally advanced NPC. The analysis of EDNRA/H323H polymorphism may help identify patient subgroups at high risk for poor disease outcome.

Topics: Adolescent; Adult; Aged; Biomarkers, Tumor; Carcinoma; Cell Line, Tumor; Cell Proliferation; Endothelin-1; Female; Gene Frequency; Genotype; Humans; Kaplan-Meier Estimate; Male; Middle Aged; Multivariate Analysis; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Outcome Assessment, Health Care; Polymorphism, Single Nucleotide; Prognosis; Proportional Hazards Models; Receptor, Endothelin A; Risk Factors; Young Adult

The peptide endothelin-1 (ET1), which was originally identified as a vasoconstrictor, has emerged as a critical regulator of a number of painful conditions, including inflammatory pain and tumor-associated pain. There is considerable pharmacological evidence supporting a role for endothelin A receptors (ET(A)) in mediating ET1-induced pro-algesic functions. ET(A) receptors are expressed in small-diameter nociceptive neurons, but also found in a variety of other cell types in peripheral tissues, including immune cells, keratinocytes, endothelial cells, which have the potential to modulate nociception. To elucidate the functional contribution of ET(A) receptors expressed in sensory neurons towards the functions of the ET1 axis in pathological pain states, we undertook a conditional gene deletion approach to selectively deplete expression of ET(A) in sensory nerves, preserving expression in non-neural peripheral tissues; the expression of ET(B) remained unchanged. Behavioural and pharmacological experiments showed that only late nociceptive hypersensitivity caused by ET1 is abrogated upon a loss of ET(A) receptors on nociceptors and further suggest that ET1-induced early nociceptive hypersensitivity involves activation of ET(A) as well as ET(B) receptors in non-neural peripheral cells. Furthermore, in the context of alleviation of cancer pain and chronic inflammatory pain by ET(A) receptor antagonists, we observed in corresponding mouse models that the contribution of ET(A) receptors expressed in nociceptors is most significant. These results help understand the role of ET(A) receptors in complex biological processes and peripheral cell-cell interactions involved in inflammatory and tumor-associated pain.

Topics: Analysis of Variance; Animals; Carcinoma; Disease Models, Animal; Endothelin A Receptor Antagonists; Endothelin-1; Ganglia, Spinal; Hyperalgesia; Lung Neoplasms; Mice; Mice, Inbred C57BL; Mice, Transgenic; NAV1.8 Voltage-Gated Sodium Channel; Nociceptors; Pain; Pain Measurement; Peptides, Cyclic; Receptor, Endothelin A; RNA, Messenger; Sensory Receptor Cells; Sodium Channels; Time Factors; Tubulin

Pancreatic stellate cells (PSC) play a key role in pancreatic fibrosis. Activation of PSC occurs in response to pro-fibrogenic stimuli and is maintained by autocrine loops of mediators, such as endothelin (ET)-1. Here, we have evaluated effects of the dual ET receptor antagonist bosentan in models of pancreatic fibrogenesis and cancer. Cell culture studies revealed that PSC and DSL6A pancreatic cancer cells expressed both ET-1 and ET receptors. Bosentan efficiently inhibited proliferation of both cell types and collagen synthesis in PSC. Expression of the myofibroblastic marker alpha-smooth muscle actin, connective tissue growth factor, and ET-1 itself in PSC was reduced, while expression of matrix metalloproteinase-9 was enhanced. Like PSC, DSL6A cells secrete less ET-1 when cultured with bosentan. In a rat model of pancreatic fibrosis, chronic pancreatitis induced by dibutyltin dichloride, a tendency towards a diminished disease progression was observed in a subgroup of rats with less severe disease. Together, our results indicate that bosentan exerts antifibrotic and antitumor effects in vitro. Its efficiency in vivo warrants further investigation.

Topics: Animals; Bosentan; Carcinoma; Cell Line, Tumor; Coculture Techniques; Collagen; Endothelin Receptor Antagonists; Endothelin-1; Fibrosis; Organotin Compounds; Pancreas; Pancreatic Neoplasms; Pancreatitis, Chronic; Rats; Receptors, Endothelin; Sulfonamides

Topics: Biomarkers, Tumor; Carcinoma; Cell Proliferation; Disease-Free Survival; Endothelin-1; Humans; Ki-67 Antigen; Mutation; Neoplasm Invasiveness; Neoplasm Staging; Prognosis; Receptor, Endothelin A; Receptor, Endothelin B; Receptor, Fibroblast Growth Factor, Type 3; Risk Assessment; Risk Factors; Time Factors; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms; Urothelium

The endothelin (ET) axis plays a role in cancer biology and plays a potential role as a target for molecular therapy in urogenital tumours. Alterations of several proteins of the ET axis were detected in invasive bladder cancer.. To examine the potential role of the expression of ET axis proteins compared to other prognostic parameters (kinase inhibitor 67 [Ki-67], tumour protein 53 [TP53], and fibroblast growth factor receptor 3 gene [FGFR3] mutations) in noninvasive and invasive bladder cancer.. Tissue microarrays from 154 consecutive patients with pTa-pT2 urothelial bladder cancer were immunohistochemically stained for endothelin 1 (ET-1), endothelin A and B receptors (ET(A)R, ET(B)R), TP53, and Ki-67. FGFR3 mutations were detected by SNaPshot analysis.. The results were correlated with clinicopathologic parameters and disease-specific survival, overall survival, and recurrence-free survival.. Proteins of the ET axis were frequently expressed in bladder cancer (ET-1 in 62% of tumours, ET(A)R in 93% of tumours, and ET(B)R in 84% of tumours). ET-1 expression was strongly correlated with tumour stage (p=0.015), histologic grade (p=0.008), and low proliferation status (p=0.003). ET(A)R immunostaining was only associated with low proliferation status (p=0.015). Kaplan-Meier survival analysis showed a significantly longer overall survival for patients with ET-1-expressing tumours (p=0.007). A significantly longer disease-free survival was found in patients with ET(A)R-expressing tumours (p=0.040), whereas ET(B)R expression was significantly correlated to a longer disease-free survival only in subgroups of patients with multifocal tumours (p=0.031), low proliferation index (Ki-67 ≤10; p=0.050), low TP53 expression (≤10; p=0.018), and tumours with an FGFR3 mutation (p=0.026). In the global model for recurrence-free survival, only high-grade (p=0.048) and negative ET(A)R immunoreactivity (p=0.048) were correlated with poor prognosis.. In addition to other factors, particularly age at diagnosis and growth pattern, lack of ET-1 expression may be an independent negative prognostic factor for the overall-survival probability of bladder cancer patients. Lack of ET(A)R expression may be an independent negative marker for recurrence-free survival.

Topics: Aged; Biomarkers, Tumor; Carcinoma; Cell Proliferation; Chi-Square Distribution; Disease-Free Survival; Endothelin-1; Female; Humans; Immunohistochemistry; Kaplan-Meier Estimate; Ki-67 Antigen; Male; Mutation; Neoplasm Invasiveness; Neoplasm Staging; Prognosis; Proportional Hazards Models; Receptor, Endothelin A; Receptor, Endothelin B; Receptor, Fibroblast Growth Factor, Type 3; Risk Assessment; Risk Factors; Time Factors; Tissue Array Analysis; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms; Urothelium

Topics: Biomarkers, Tumor; Carcinoma; Cell Proliferation; Disease-Free Survival; Endothelin-1; Humans; Ki-67 Antigen; Mutation; Neoplasm Invasiveness; Neoplasm Staging; Prognosis; Receptor, Endothelin A; Receptor, Endothelin B; Receptor, Fibroblast Growth Factor, Type 3; Risk Assessment; Risk Factors; Time Factors; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms; Urothelium

Endothelin-1 (ET-1) is a potent vasoactive peptide and a hypoxia-inducible angiogenic growth factor associated with the development and spread of solid tumors. The clinical significance of plasma big ET-1 in patients with advanced-stage nasopharyngeal carcinoma (NPC) is not known.. Pretreatment plasma big ET-1 levels were measured in 62 patients with advanced-stage NPC using a sandwich enzyme-linked immunoassay and compared with the levels from a control group (n = 19 participants).. The median pretreatment plasma big ET-1 level in patients with advanced-stage NPC was 4.6 pg/mL (range, 1.9-15.2 pg/mL) and was significantly elevated compared with median plasma big ET-1 levels in healthy controls, 2.6 pg/mL (1.6-4.5 pg/mL) (P < .001). Using the upper limit (4.5 pg/mL) of control subjects as the cut-off value, plasma big ET-1 was < or = 4.5 pg/mL in 29 (46.8%) patients and > 4.5 pg/mL in 33 (53.2%) patients. A pretreatment plasma big ET-1 level > 4.5 pg/mL was associated with a significantly poorer 2-year distant metastasis-free survival rate (56.7% vs. 81.1%, P = .031). Multivariate analysis showed that N classification (hazard ratio [HR], 2.416; 95% confidence interval [CI], 1.071-5.447; P = .034) and pretreatment plasma big ET-1 level (HR, 3.151; 95% CI, 1.099-9.028, P = .033) were independent significant prognostic factors for posttreatment distant failure in patients with advanced-stage NPC.. Pretreatment plasma big ET-1 levels may be useful in predicting posttreatment distant failure in patients with advanced-stage NPC.

Topics: Adult; Biomarkers, Tumor; Carcinoma; Case-Control Studies; Endothelin-1; Enzyme-Linked Immunosorbent Assay; Female; Humans; Male; Middle Aged; Multivariate Analysis; Nasopharyngeal Neoplasms; Neoplasm Metastasis; Neoplasm Staging; Prognosis; Survival Analysis; Treatment Outcome

Endothelin-1 (ET-1) is overexpressed in breast carcinomas and influences via its receptors (ETAR and ETBR) transformation, differentiation and growth processes in the human breast, but little is known about the ET expression in breast cancer precursors. On this basis we evaluated the expression of ET-1, ETAR and ETBR in a series of breast carcinomas, ductal (DCIS) and lobular carcinoma in situ (LCIS) and normal breast tissue by immunohistochemical (IH) methods. IH staining of ET-1, ETAR and ETBR was performed in 88 invasive breast carcinomas, with adjacent carcinoma in situ and concomitant normal breast tissue. Moderate or strong cytoplasmic immunostaining was observed for ET-1 in 33.3%, for ETAR in 45.3% and for ETBR in 55.7% of invasive breast carcinomas. Comparative analysis of invasive cancer (CA), concomitant carcinoma in situ (CIS) and normal breast epithelium (NBE) revealed a stepwise increase of ET-1 and ETAR expression in the sequence NBE < CIS < CA. ETBR expression tended to be slightly higher in CIS than in CA (NBE versus CIS and NBE versus CA, for ETAR and ETBR, p<0.001, respectively; NBE versus CA for ET-1, p=0.035). Our data suggest that the expression of ET-1, ETAR and ETBR correlates with the acquisition of malignant potential and may be used as a prognostic indicator of aggressive behaviour and invasive potential of premalignant breast lesions.

Topics: Biomarkers, Tumor; Breast Neoplasms; Carcinoma; Carcinoma in Situ; Carcinoma, Intraductal, Noninfiltrating; Carcinoma, Lobular; Endothelin-1; Female; Humans; Immunochemistry; Receptor, Endothelin A; Receptor, Endothelin B; Retrospective Studies

Endothelin-1 (ET-1) and its receptors (ET(A)R and ET(B)R), referred to as the endothelin (ET) axis, are overexpressed in breast carcinomas, and influence tumorigenesis and tumor progression by various mechanisms, including angiogenesis. The objective of the study was to clarify if expression of the ET axis participates in angiogenesis of breast carcinoma. We analyzed expression of ET-1, ET(A)R, ET(B)R, and vascular endothelial growth factor (VEGF) immunohistochemically in 600 tissue array specimens from 200 paraffin-embedded breast carcinomas performing tissue microarray technology. Microvessel density (MVD) was determined by counting microvessels (identified by factor VIII) in each core specimen.. Moderate or strong immunostaining was observed for ET-1 in 25.4%, for ET(A)R in 43.7%, and for ET(B)R in 22.2% of breast carcinomas. Of all cases, 44.7% showed significant expression of VEGF. MVD varied between different tumor specimens (range, 0-80; median, 17). We observed a statistically significant correlation between MVD and ET expression status with higher MVD in ET-positive tumors. Moreover, expression of VEGF was found more frequently in tumors with overexpression of the ET axis (each P < 0.001). Staining of VEGF was correlated positively with MVD CONCLUSIONS: These results indicate that increased ET-1, ET(A)R, and ET(B)R expression is associated with increased VEGF expression and higher vascularity of breast carcinomas and, thus, could be involved in the regulation of angiogenesis in breast cancer. Our findings provide evidence that the expression pattern of the ET-axis and in particular of ET(A)R may have clinical relevance in future antiangiogenic targeted therapies for breast cancer.

Topics: Breast Neoplasms; Carcinoma; Cell Line, Tumor; Disease Progression; Endothelin-1; Factor VIII; Humans; Immunohistochemistry; Microcirculation; Neoplasm Metastasis; Neovascularization, Pathologic; Platelet Endothelial Cell Adhesion Molecule-1; Receptor, Endothelin A; Receptor, Endothelin B; Tissue Distribution; Vascular Endothelial Growth Factor A

The vasodilator hydralazine, used clinically in cardiovascular therapy, relaxes arterial smooth muscle by inhibiting accumulation of intracellular free Ca2+ via an unidentified primary target. Collagen prolyl hydroxylase is a known target of hydralazine. We therefore investigated whether inhibition of other members of this enzyme family, namely the hypoxia-inducible factor (HIF)-regulating O2-dependent prolyl hydroxylase domain (PHD) enzymes, could represent a novel mechanism of action. Hydralazine induced rapid and transient expression of HIF-1alpha and downstream targets of HIF (endothelin-1, adrenomedullin, haem oxygenase 1, and vascular endothelial growth factor [VEGF]) in endothelial and smooth muscle cells and induced endothelial cell-specific proliferation. Hydralazine dose-dependently inhibited PHD activity and induced nonhydroxylated HIF-1alpha, evidence for HIF stabilization specifically by inhibition of PHD enzyme activity. In vivo, hydralazine induced HIF-1alpha and VEGF protein in tissue extracts and elevated plasma VEGF levels. In sponge angiogenesis assays, hydralazine increased stromal cell infiltration and blood vessel density versus control animals. Thus, hydralazine activates the HIF pathway through inhibition of PHD activity and initiates a pro-angiogenic phenotype. This represents a novel mechanism of action for hydralazine and presents HIF as a potential target for treatment of ischemic disease.

Topics: Adrenomedullin; Angiogenesis Inducing Agents; Animals; Breast Neoplasms; Carcinoma; Carcinoma, Renal Cell; Cell Hypoxia; Cell Line, Tumor; Cells, Cultured; DNA-Binding Proteins; Dose-Response Relationship, Drug; Endothelial Cells; Endothelin-1; Enzyme Inhibitors; Gene Expression Regulation; Heme Oxygenase (Decyclizing); Heme Oxygenase-1; Humans; Hydralazine; Hypoxia-Inducible Factor 1; Hypoxia-Inducible Factor 1, alpha Subunit; Implants, Experimental; Kidney Neoplasms; Membrane Proteins; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Nude; Myocytes, Smooth Muscle; Neovascularization, Physiologic; Nuclear Proteins; Peptides; Procollagen-Proline Dioxygenase; Transcription Factors; Vascular Endothelial Growth Factor A; Vasodilator Agents

Endothelin-1 (ET-1) is overexpressed in ovarian carcinomas and acts, via ET(A) receptors (ET(A)R), as an autocrine growth factor. In this study we investigate the role of ET-1 in the neovascularization of ovarian carcinoma. Archival specimens of primary (n = 40) and metastatic (n = 8) ovarian tumors were examined by immunohistochemistry for angiogenic factor and receptor expression and for microvessel density using antibodies against CD31, ET-1, vascular endothelial growth factor (VEGF), and their receptors. ET-1 expression correlated with neovascularization and with VEGF expression. The localization of functional ET(A)R and ET(A)R mRNA expression, as detected by autoradiography and in situ hybridization, was evident in tumors and in intratumoral vessels, whereas ET(B)R were expressed mainly in endothelial cells. High levels of ET-1 were detected in the majority of ascitic fluids of patients with ovarian carcinoma and significantly correlated with VEGF ascitic concentration. Furthermore ET-1, through ET(A)R, stimulated VEGF production in an ovarian carcinoma cell line, OVCA 433, by an extent comparable to hypoxia. Finally, conditioned media from OVCA 433 as well as ascitic fluids caused an increase in endothelial cell migration and the ET-1 receptor blockade significantly inhibited this angiogenic response. These findings indicate that ET-1 could modulate tumor angiogenesis, acting directly and in part through VEGF.

Topics: Adenocarcinoma; Adult; Aged; Ascitic Fluid; Blood Vessels; Carcinoma; Cell Movement; Endothelial Growth Factors; Endothelin-1; Endothelium, Vascular; Female; Humans; Lymphokines; Middle Aged; Neovascularization, Pathologic; Ovarian Neoplasms; Receptor, Endothelin A; Receptor, Endothelin B; Receptors, Endothelin; Tumor Cells, Cultured; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Choroid plexus carcinoma is a rare neoplasm derived from the epithelium of the choroid plexus. The production and secretion of endothelin-1 (ET-1) by cultured human choroid plexus carcinoma cells were studied by radioimmunoassay and Northern blot analysis. Immunoreactive (IR)-ET was detected in the culture medium (2.78 +/- 0.12 fmol/10(5) cells/24 h; n = 5; mean +/- SEM) but not in the unconditioned medium. Reverse-phase high-performance liquid chromatography of the extract of the culture medium showed a single peak eluting in the position of ET-1. Treatment with tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta) or a combination of interferon-gamma (IFN-gamma), TNF-alpha, and IL-1 beta caused significant increases in the IR-ET levels in the culture medium. Northern blot analysis of total RNA showed the expression of ET-1 mRNA in choroid plexus carcinoma cells. The expression levels of ET-1 mRNA were increased by treatment with a combination of IFN-gamma, TNF-alpha, and IL-1 beta. The present study has shown the production and secretion of ET-1 by cultured human choroid plexus carcinoma cells and suggests the possibility that ET-1 formation is related to the pathophysiology of this tumor.

Topics: Blotting, Northern; Calcitonin Gene-Related Peptide; Carcinoma; Choroid Plexus Neoplasms; Chromatography, High Pressure Liquid; Cytokines; Endothelin-1; Humans; Neuropeptide Y; Radioimmunoassay; Tumor Cells, Cultured

To elucidate the presence and distribution of Endothelin-1 (ET-1) in tissues of human salivary glands, we performed an immunohistochemical analysis of ET-1 in 15 normal salivary glands, 8 adenolymphomas, 13 pleomorphic adenomas and 5 carcinomas, using the mouse monoclonal antibody against human ET-1. In normal glands, immunoreactivity for ET-1 was observed in the striated duct cells. In adenolymphomas, the columnar cells of the granular epithelium showed strong intracytoplasmic immunoreactivity. In carcinomas, moderate or strong immunoreactivity was observed in the tumor cells, whereas in pleomorphic adenomas, weak immunoreactivity was observed. A good relation was detected between the size of pleomorphic adenoma and ET-1 immunoreactivity, as well as between the duration of tumor in carcinoma and ET-1 immunoreactivity. The presence and distribution of ET-1 in salivary glands and salivary gland tumors suggests a possible role for ET-1 in the regulation of electrolytes and water transport in salivary glands, and as a growth-promoting factor for tumors.

Topics: Adenolymphoma; Adenoma, Pleomorphic; Aged; Carcinoma; Endothelin-1; Female; Humans; Immunohistochemistry; Male; Middle Aged; Salivary Gland Neoplasms; Salivary Glands

Using electron immunocytochemistry, blood vessels in the normal rat liver and in 2 different animal models of liver metastases: (1) Hooded Lister rat with MC28 tumour, a sarcoma, and (2) nude rat with HT29 tumour, a carcinoma, were investigated for the presence of endothelin-1. In the normal livers, small subpopulations of vascular endothelial cells displayed discrete immunoreactivity for endothelin-1. In the livers with malignant tumours, there was a substantial increase in endothelin-1-immunoreactive endothelial cells in vessels located at the tumour periphery. In the controls, antibody to endothelin-1 also labelled sporadically some fibroblast/fibroblast-like cells associated with the blood vessels. In contrast, intense immunoreactivity for endothelin-1 was frequently associated with the tumour cells and/or fibroblast cells in both types of tumour examined.

Topics: Animals; Carcinoma; Endothelin-1; Endothelium, Vascular; Fibroblasts; Liver; Liver Neoplasms, Experimental; Male; Microscopy, Immunoelectron; Rats; Rats, Inbred Strains; Rats, Nude; Sarcoma, Experimental