endothelin-1 and Urinary-Bladder-Neoplasms

RhoGDI2 is a guanine nucleotide dissociation inhibitor (GDI) specific for the Rho family of small GTPases that plays dual opposite roles in tumor progression, being both a promoter in tissues such as breast and a metastasis suppressor in tissues such as the bladder. Despite a clear role for this protein in modulating the invasive and metastatic process, the mechanisms through which RhoGDI2 executes these functions remain unclear. This review will highlight the current state of our knowledge regarding how RhoGDI2 functions in metastasis with a focus on bladder cancer and will also seek to highlight other potential underappreciated avenues through which this protein may affect cancer cell behavior.

Topics: Animals; Apoptosis; Endothelin-1; GTP Phosphohydrolases; Humans; Neoplasm Metastasis; Protein Kinase C-alpha; rho Guanine Nucleotide Dissociation Inhibitor beta; Urinary Bladder Neoplasms

The endothelin system comprises the three peptide hormones endothelin (ET)-1, -2, -3, their G protein-coupled receptors, endothelin-A-receptor (ET(A)R) and endothelin-B-receptor (ET(B)R), and the enzymes of endothelin biosynthesis and degradation. In the past two decades, an impressive amount of data has been accumulated investigating the role of the endothelin system in a variety of malignancies. In many cancers, ET-1/ET(A)R interaction induces proliferation, angiogenesis, antiapoptosis and resistance to chemotherapy. Data indicate a pivotal role of the endothelin system in tumorigenesis, local progression and metastasis. Subsequently, novel drugs have been designed inhibiting ET-1 biosynthesis or ET(A)R interaction. A wide range of preclinical data is available on the role of ET(A)R antagonists in gynecological, urological and breast cancers providing evidence for their antiangiogenic, proapoptotic and growth inhibitory effects. Of particular interest is the anti-invasive and antimetastatic efficacy of ET(A)R antagonists and synergism when co-administered with established cancer therapies. Data indicate a future role of ET(A)R antagonists in oncologic therapies.

Topics: Animals; Antineoplastic Agents; Aspartic Acid Endopeptidases; Breast Neoplasms; Endometrial Neoplasms; Endothelin Receptor Antagonists; Endothelin-1; Endothelin-Converting Enzymes; Female; Humans; Kidney Neoplasms; Male; Metalloendopeptidases; Ovarian Neoplasms; Prostatic Neoplasms; Receptors, Endothelin; Urinary Bladder Neoplasms; Urogenital Neoplasms; Uterine Cervical Neoplasms

To conduct a prospective study of the potential prognostic role of endothelin-1 (ET-1) in a cohort of primary high-grade non-muscle-invasive urothelial bladder cancer patients, who were treated with adjuvant intravesical Bacillus Calmette-Guérin (BCG).. Patients with primary high-grade nonmuscle- invasive urothelial bladder cancer, who received postoperatively induction and maintenance BCG therapy, were prospectively included. Recurrence and progression were histologically proven. Immunohistochemical staining for ET-1 was assessed. Epidemiological, pathological and clinical parameters as well as the expression of ET-1 in tumor specimens were statistically analyzed for recurrence, progression, recurrence-free survival (RFS) and progression-free survival (PFS).. ET-1 associates significantly with recurrence (p = 0.000), progression (p = 0.000), RFS (p = 0.000) and PFS (p = 0.000). The patient's age is also significant for recurrence (p = 0.003, OR = 1.273 95% CI: 1.086-1.492) and RFS (p = 0.013).. ET-1 seems to deteriorate prognosis in patients suffering from primary high-grade non-muscle-invasive urothelial bladder cancer, who are treated with adjuvant BCG instillations. Furthermore, the patient's age associates with an increased likelihood for recurrence.

Topics: Adjuvants, Immunologic; BCG Vaccine; Carcinoma, Transitional Cell; Endothelin-1; Humans; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Prognosis; Prospective Studies; Urinary Bladder Neoplasms

The monitoring of vascular-targeted therapies using magnetic resonance imaging, computed tomography or ultrasound is limited by their insufficient spatial resolution. Here, by taking advantage of the intrinsic optical properties of haemoglobin, we show that raster-scanning optoacoustic mesoscopy (RSOM) provides high-resolution images of the tumour vasculature and of the surrounding tissue, and that the detection of a wide range of ultrasound bandwidths enables the distinction of vessels of differing size, providing detailed insights into the vascular responses to vascular-targeted therapy. Using RSOM to examine the responses to vascular-targeted photodynamic therapy in mice with subcutaneous xenografts, we observed a substantial and immediate occlusion of the tumour vessels followed by haemorrhage within the tissue and the eventual collapse of the entire vasculature. Using dual-wavelength RSOM, which distinguishes oxyhaemoglobin from deoxyhaemoglobin, we observed an increase in oxygenation of the entire tumour volume immediately after the application of the therapy, and a second wave of oxygen reperfusion approximately 24 h thereafter. We also show that RSOM enables the quantification of differences in neoangiogenesis that predict treatment efficacy.

Topics: Animals; Brain; Cerebral Ventricle Neoplasms; Colonic Neoplasms; Craniotomy; Diagnostic Imaging; Disease Models, Animal; Endothelin-1; Epinephrine; Female; Heterografts; Image Processing, Computer-Assisted; Imaging, Three-Dimensional; Lasers; Mice; Mice, Inbred BALB C; Neovascularization, Pathologic; Oxygen; Photoacoustic Techniques; Ultrasonography; Urinary Bladder Neoplasms; Vascular Neoplasms; Vasoconstriction

Bladder cancer (BC) is the second most common malignancy of the genitourinary system, characterized by the highest recurrence rate of all cancers. Treatment options are limited; thus a thorough understanding of the underlying molecular mechanisms is needed to guide the discovery of novel therapeutic targets. Profilins are actin binding proteins with attributed pleiotropic functions to cytoskeletal remodeling, cell adhesion, motility, even transcriptional regulation, not fully characterized yet. Earlier studies from our laboratory revealed that decreased tissue levels of Profilin-1 (PFN1) are correlated with BC progression to muscle invasive disease. Herein, we describe a comprehensive analysis of PFN1 silencing via shRNA, in vitro (by employing T24M cells) and in vivo [(with T24M xenografts in non-obese diabetic severe combined immunodeficient mice (NOD/SCID) mice]. A combination of phenotypic and molecular assays, including migration, proliferation, adhesion assays, flow cytometry and total mRNA sequencing, as well as immunohistochemistry for investigation of selected findings in human specimens were applied. A decrease in BC cell adhesion and tumor growth in vivo following PFN downregulation are observed, likely associated with the concomitant downregulation of Fibronectin receptor, Endothelin-1, and Actin polymerization. A decrease in the levels of multiple key members of the non-canonical Wnt/Ca2+ signaling pathway is also detected following PFN1 suppression, providing the groundwork for future studies, addressing the specific role of PFN1 in Ca2+ signaling, particularly in the muscle invasive disease.

Topics: Actins; Animals; Calcium; Cell Adhesion; Cell Line, Tumor; Cell Movement; Cell Proliferation; Disease Progression; Down-Regulation; Endothelin-1; Flow Cytometry; Humans; Immunohistochemistry; Integrin alpha5beta1; Male; Mice; Mice, Inbred NOD; Mice, SCID; Muscle Neoplasms; Profilins; Protein Multimerization; RNA Interference; RNA, Small Interfering; Urinary Bladder; Urinary Bladder Neoplasms; Wnt Signaling Pathway; Xenograft Model Antitumor Assays

Endothelin-1 (ET-1) is a multifunctional peptide exerting its effects via receptors A and B. ET-1 and its receptors, endothelin axis (ET axis), play a promoting role in cancer biology. Alterations of proteins of ET axis have been detected in non-metastatic muscle-invasive bladder cancer (NMMIBC). The objective of this study is to investigate the potential role of ET-1 tumor expression as a biomarker of prognosis, compared to other prognostic parameters (epidemiologic and pathologic), in NMMIBC. We prospectively included 40 consecutive, primary, high-grade NMMIBC patients. Tumor specimens after initial transurethral resection were stained immunohistochemically for ET-1. The ET-1 evaluation of expression was based on staining intensity (SI) of ET-1. SI was classified according to an arbitrary four-tiered scale (negative = 0, mild = 1, moderate = 2, strong = 3). Epidemiologic and pathologic parameters were analyzed, using univariate and multivariate statistics, for disease progression, progression-free survival (PFS), and overall survival (OS). ET-1 overexpression (SI = 3) was the unique parameter which associated significantly, both in univariate (log-rank test, p = 0.033) and multivariate (Cox regression analysis, p = 0.045, HR = 4.849, 95 % CI: 1.039-22.624) analysis, with an increased hazard ratio of progression. ET-1 overexpression (SI = 3) was also the unique parameter which associated, marginally significantly in univariate analysis (log-rank test, p = 0.056) and highly significantly in multivariate analysis (Cox regression analysis, p = 0.005, HR = 7.001, 95 % CI: 1.782-27.501), with an increased hazard ratio of death. Overexpression of ET-1 may be a potential biomarker of unfavorable prognosis in NMMIBC patients.

Topics: Adult; Aged; Biomarkers, Tumor; Disease-Free Survival; Endothelin-1; Female; Gene Expression Regulation, Neoplastic; Humans; Kaplan-Meier Estimate; Male; Middle Aged; Muscle Neoplasms; Muscles; Neoplasm Invasiveness; Prognosis; Urinary Bladder Neoplasms

Many patients with advanced bladder cancer develop lethal metastases to the lung. The vasoconstricting protein endothelin-1 (ET-1) has been implicated in this process, although the mechanism(s) by which it promotes metastasis remains unclear. Here, we have evaluated whether tumor ET-1 expression can serve as a biomarker for lung metastasis and whether it is required for metastatic disease. Evaluation of ET-1 mRNA and protein expression in four patient cohorts revealed that levels of ET-1 are higher in patients with muscle-invasive bladder cancers, which are associated with higher incidence of metastasis, and that high ET-1 levels are associated with decreased disease-specific survival. Consistent with its proinflammatory activity, we found that tumor-derived ET-1 acts through endothelin-1 receptor A (ETAR) to enhance migration and invasion of both tumor cells and macrophages and induces expression of inflammatory cytokines and proteases. Using human and mouse cancer cells depleted of ET-1 and pharmacologic blockade of ET receptors in lung metastasis models, we found that tumor ET-1 expression and ETAR activity are necessary for metastatic lung colonization and that this process is preceded by and dependent on macrophage infiltration of the lung. In contrast, tumor ET-1 expression and ETAR activity appeared less important in established primary or metastatic tumor growth. These findings strongly suggest that ETAR inhibitors might be more effective as adjuvant therapeutic agents than as initial treatment for advanced primary or metastatic disease.

Topics: Animals; Base Sequence; Biomarkers, Tumor; Cell Line, Tumor; Cytokines; Endothelin A Receptor Antagonists; Endothelin-1; Female; Gene Expression; Gene Knockdown Techniques; Humans; Inflammation Mediators; Lung Neoplasms; Macrophages; Mice; Mice, Nude; Neoplasm Invasiveness; Neoplasm Transplantation; Receptor, Endothelin A; RNA, Messenger; RNA, Neoplasm; RNA, Small Interfering; Transplantation, Heterologous; Urinary Bladder Neoplasms

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

The endothelin axis consists of endothelin-1 (ET-1) and its two receptors, ET(A)- and ET(B)-receptor (ET(A)-R and ET(B)-R). In several tumor entities, the ET(A)-R plays a significant role as a drug target. In our study, we investigated whether inhibition of ET(A)-R with atrasentan leads to an antitumor effect in urinary bladder carcinoma as well.. Twenty nude mice with thymic aplasia were subcutaneously administered 2 x 10(6) KU-19-19 bladder cancer cells in the right flank. Starting on the 22nd day after the injection, ten animals were treated with atrasentan (2.5 mg/kg BW intraperitoneally), and another ten animals were treated with placebo. During treatment, absolute tumor growth and relative growth rate over time were determined. After the end of treatment, the mitosis and necrosis rates, microvessel density, and receptor density in the tumor tissue were analyzed by immunohistochemistry. In addition, the expression intensities of ET-1, ET(A)-R, and ET(B)-R were evaluated semiquantitatively and compared between the groups.. No significant differences between the active-treatment and placebo groups were detected, either with respect to absolute tumor growth (P = 0.333) or mitosis rate (P = 0.217). In the analysis of the necrosis rate and receptor density for ET(A)-R, a trend toward higher values in the active-treatment group (mean necrosis rate = 63.67%, receptor density: 1.417) than in the placebo group (mean necrosis rate = 46.25%, receptor density: 1.270) was found; however, neither difference was statistically significant (P = 0.08 and 0.219, respectively).. ET(A)-R blockade with atrasentan in a bladder cancer xenograft model shows no significant antitumor effect.

Topics: Animals; Atrasentan; Disease Models, Animal; Endothelin A Receptor Antagonists; Endothelin B Receptor Antagonists; Endothelin-1; Female; Humans; Immunoenzyme Techniques; Mice; Mice, Nude; Mitosis; Necrosis; Pyrrolidines; Receptor, Endothelin A; Receptor, Endothelin B; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Thymus Gland; Urinary Bladder Neoplasms; Xenograft Model Antitumor Assays

Endothelin-1 (ET-1) and its receptors, entothelin-A (ETAR) and endothelin-B (ETBR), commonly referred to as the endothelin (ET)-axis, are involved in tumor biology and growth. We investigated the effects of the ET-axis on microvessel density (MVD) and the clinicopathological parameters of patients with invasive bladder cancer. Paraffin tumor sections of 120 patients who had undergone radical cystectomy were assessed immunohistochemically using mono- and polyclonal antibodies for ET-1, ETAR, ETBR and CD34 (MVD). Staining intensities were analyzed semiquantitatively and the MVD was calculated as vessels per field. The results were correlated with various pathological and clinical factors, as well as with disease-free and overall survival. Transitional cell carcinomas (MVD=23.7) were better vascularized than squamous cell carcinomas (MVD=17.8, p=0.04). Organ-confined tumors (MVD=32.2) were better vascularized than T3- and T4-tumors (MVD=21.2, p=0.02) and ET-1 was overexpressed in this subgroup (p=0.027). Patients with metastatic regional lymph nodes (MVD=20.9) tended to have less MVD than patients without regional lymph node metastases (MVD=24.1) (p=0.15). The account of MVD did not reveal any significant differences in disease-free or overall survival. Organ-confined tumors and ET-1 overexpression are associated with upregulated microvessel density. These results suggest that MVD and ET-1 could be considered good prognostic factors.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Cystectomy; Endothelin-1; Female; Humans; Immunoenzyme Techniques; Lymph Nodes; Male; Middle Aged; Neoplasm Staging; Neovascularization, Pathologic; Prognosis; Receptor, Endothelin A; Receptor, Endothelin B; Survival Rate; Urinary Bladder Neoplasms

Overexpression of endothelin-1, endothelin-A- and endothelin-B-receptors has been shown in various human tumors. To assess the role of the ET-axis in bladder cancer, we analyzed its expression in tumor specimens and bladder cancer cell lines. Samples were obtained by radical cystectomy at two urologic institutions. ET-axis expression was investigated by reverse-transcription polymerase chain reaction (RT-PCR) (n=22) and immunohistochemistry (n=42). Additionally, four bladder cancer cell lines were analyzed. RT-PCR analysis for the ET-axis showed positive signals in the majority of cDNA probes. Signals for endothelin-1 (ET-1), endothelin-A-receptor (ET(A)R) and endothelin-B-receptor (ET(B)R), as identified semiquantitatively and by densitometry, were found in 22, 22 and 15 of 22 cases, respectively. Immunohistochemistry revealed expression of ET-1, ET(A)- and ET(B)-receptor in 18, 29 and 37 of the 42 cases, respectively, whereas normal urothelium was negative. All cell lines expressed ET-1, and all but the RT-112 cell line produced ETAR, whereas no cell line expressed ET(B)R. The identification of the ET-axis at the mRNA and protein level in the majority of bladder tumor samples suggests a role in the carcinogenesis of bladder cancer. Further studies on regulation of the ET-axis and the future use of selective ET(A)-receptor inhibitors for targeted molecular therapy in bladder cancer are in progress.

Topics: Adult; Aged; Carcinoma, Transitional Cell; Endothelin-1; Female; Humans; Middle Aged; Receptor, Endothelin A; Receptor, Endothelin B; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Cells, Cultured; Urinary Bladder Neoplasms

Endothelin-1 (ET-1) and its receptors ET(A)R and ET(B)R, referred to as the Endothelin-axis, play an emerging role in cancer. We examined the ET-axis immunohistochemically in invasive bladder cancer.. Tumor specimens from 157 patients after cystectomy were stained immunohistochemically for ET-1, ET(A)R and ET(B)R. After semiquantitative analysis the staining results were correlated with clinicopathological parameters and survival rates.. Overexpression of ET-1, ET(A)R and ET(B)R was identified in 26.8%, 58.8% and 76.9% of cases, respectively. No association with TNM staging and histologic grading was found. However, patients with ET(B)R expression tended to have organ-confined tumors (p=0.16) and no vascular invasion (p=0.09), the latter being statistically significant in the subgroup of G3 tumors (p=0.02). ET(B)R overexpression was associated with favorable disease-free survival (p=0.04).. The ET-axis is overexpressed in bladder cancer, ET(B)R predominating in this entity and being associated with a more favorable prognosis. Further studies are warranted to elucidate the role of the ET-axis as a molecular target in bladder cancer.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Cystectomy; Disease-Free Survival; Endothelin-1; Female; Follow-Up Studies; Humans; Immunohistochemistry; Male; Middle Aged; Neoplasm Staging; Prognosis; Receptor, Endothelin A; Receptor, Endothelin B; Survival Rate; Time Factors; Urinary Bladder Neoplasms

Half of patients treated for locally advanced bladder cancer relapse with often fatal metastatic disease to the lung. We have recently shown that reduced expression of the GDP dissociation inhibitor, RhoGDI2, is associated with decreased survival of patients with advanced bladder cancer. However, the effectors by which RhoGDI2 affects metastasis are unknown. Here we use DNA microarrays to identify genes suppressed by RhoGDI2 reconstitution in lung metastatic bladder cancer cell lines. We identify such RNAs and focus only on those that also increase with tumor stage in human bladder cancer samples to discover only clinically relevant targets of RhoGDI2. Levels of endothelin-1 (ET-1), a potent vasoconstrictor, were affected by both RhoGDI2 reconstitution and tumor stage. To test the hypothesis that the endothelin axis is important in lung metastasis, lung metastatic bladder carcinoma cells were injected in mice treated with the endothelin receptor-specific antagonist, atrasentan, thereby blocking engagement of the up-regulated ET-1 ligand with its cognate receptor. Endothelin antagonism resulted in a dramatic reduction of lung metastases, similar to the effect of reexpressing RhoGDI2 in these metastatic cells. Taken together, these experiments show a novel approach of identifying therapeutic targets downstream of metastasis suppressor genes. The data also suggest that blockade of the ET-1 axis may prevent lung metastasis, a new therapeutic concept that warrants clinical evaluation.

Topics: Animals; Cell Line, Tumor; Down-Regulation; Endothelin-1; Gene Expression Regulation, Neoplastic; Genes, Tumor Suppressor; Guanine Nucleotide Dissociation Inhibitors; Humans; Lung Neoplasms; Mice; Mice, Nude; Neoplasm Staging; Oligonucleotide Array Sequence Analysis; rho Guanine Nucleotide Dissociation Inhibitor beta; rho-Specific Guanine Nucleotide Dissociation Inhibitors; Transfection; Tumor Suppressor Proteins; Urinary Bladder Neoplasms