endothelin-1 and Lung-Neoplasms

Topics: Alcohol Drinking; Antioxidants; Dose-Response Relationship, Drug; Endothelin-1; Female; Flavonoids; Food, Organic; Heart Diseases; Humans; Lung Neoplasms; Male; Phenols; Polyphenols; Prostatic Neoplasms; Wine

Endothelin-1 (ET-1) is a 21 amino acid peptide with diverse biological activity that has been implicated in numerous diseases. ET-1 is a potent mitogen regulator of smooth muscle tone, and inflammatory mediator that may play a key role in diseases of the airways, pulmonary circulation, and inflammatory lung diseases, both acute and chronic. This review will focus on the biology of ET-1 and its role in lung disease.

Topics: Amino Acid Sequence; Endothelin-1; Graft Rejection; Humans; Lung; Lung Diseases; Lung Neoplasms; Lung Transplantation; Molecular Sequence Data; Respiratory Distress Syndrome; Respiratory Tract Diseases; Vascular Diseases

Previous evidence has implied that obesity is an independent risk factor for developing cancer. Being closely related to obesity, type 2 diabetes mellitus provides a suitable environment for the formation and metastasis of tumors through multiple pathways. Although bariatric surgeries are effective in preventing and lowering the risk of various types of cancer, the underlying mechanisms of this effect are not clearly elucidated.. To uncover the role and effect of sleeve gastrectomy (SG) in preventing lung cancer in obese and diabetic rats.. SG was performed on obese and diabetic Wistar rats, and the postoperative transcriptional and translational alterations of the endothelin-1 (ET-1) axis in the lungs were compared to sham-operated obese and diabetic rats and age-matched healthy controls to assess the improvements in endothelial function and risk of developing lung cancer at the postoperative 4. Compared to obese and diabetic sham-operated rats, SG brought a significant reduction to body weight, food intake, and fasting blood glucose while improving oral glucose tolerance and insulin sensitivity. In addition, ameliorated levels of gene and protein expression in the ET-1 axis as well as reduced DNA damage indicated improved endothelial function and a lower risk of developing lung cancer after the surgery.. Apart from eliminating metabolic disorders, SG improves endothelial function and plays a protective role in preventing lung cancer

Topics: Animals; Bariatric Surgery; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diet, High-Fat; DNA Breaks, Double-Stranded; Endothelin-1; Endothelium; Glucose Tolerance Test; Histones; Humans; Lung; Lung Neoplasms; Male; Obesity; Phosphoproteins; Phosphorylation; Rats; Streptozocin; Weight Loss

Progression on therapy in non-small cell lung carcinoma (NSCLC) is often evaluated radiographically, however, image-based evaluation of said therapies may not distinguish disease progression due to intrinsic tumor drug resistance or inefficient tumor penetration of the drugs. Here we report that the inhibition of mutated

Topics: Animals; Antineoplastic Agents; Biological Availability; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Drug Resistance, Neoplasm; Endothelin-1; ErbB Receptors; Erlotinib Hydrochloride; Gefitinib; Humans; Lung Neoplasms; Mice; Mutation; Protein Kinase Inhibitors; Vascular Endothelial Growth Factor A; Vasoconstriction; Xenograft Model Antitumor Assays

GDF-15 is an established cardiovascular risk marker but is equally implicated in tumour biology. Elevated levels of GDF-15 have indeed been observed in distinct tumour entities. This study aimed to explore the relation of GDF-15 to other cardiac biomarkers and the general association of GDF-15 on prognosis in an unselected cohort of treatment-naïve cancer patients.. We prospectively enrolled 555 consecutive patients at time of diagnosis of malignant disease prior receiving anticancer therapy. Plasma GDF-15 concentrations were determined alongside other cardiac and routine laboratory markers. All-cause mortality was defined as primary endpoint.. GDF-15 levels were 338 ng/L (IQR:205-534) for the total cohort, and values were comparable for different tumour entities except breast cancer. Metastatic disease was characterized by higher plasma GDF-15 [435 ng/L (IQR:279-614) vs 266 ng/L (IQR:175-427), P < .001]. GDF-15 correlated positively with inflammatory status reflected by CRP, SAA and IL-6 [r = .31, P < .001, r = .23, P < .001 and r = .14, P = .002] and cardiac biomarkers as NT-proBNP, hsTnT, MR-proADM and CT-proET-1 [r = .46; r = .46; r = .59 and r = .50; P < .001 for all]. GDF-15 was significantly associated with all-cause mortality after multivariate adjustment [adj.HR for ln(GDF-15) 1.78, 95%CI:1.47-2.16, P < .001]. There was a significant interaction between solid and haematological malignancies with loss of association of GDF-15 with outcome in myelodysplastic and myeloproliferative disease.. Elevated plasma GDF-15 is associated with progressing disease severity and poor prognosis in solid tumours of treatment-naïve cancer patients. GDF-15 increase is accompanied by worsening systemic inflammation and a subclinical functional impairment of different organs including the heart. GDF-15 represents a promising target for our pathophysiologic understanding in cardio-oncology linking conditions of both cardiac and neoplastic disease.

Topics: Adrenomedullin; Aged; Breast Neoplasms; C-Reactive Protein; Cause of Death; Endothelin-1; Female; Gastrointestinal Neoplasms; Glycopeptides; Growth Differentiation Factor 15; Humans; Interleukin-6; Lung Neoplasms; Male; Middle Aged; Mortality; Myelodysplastic Syndromes; Myeloproliferative Disorders; Natriuretic Peptide, Brain; Neoplasm Metastasis; Neoplasms; Peptide Fragments; Prognosis; Proportional Hazards Models; Prospective Studies; Protein Precursors; Serum Amyloid A Protein; Troponin T

LAM is a rare low-grade metastasizing lung neoplasm. Inhibitors of mTOR improve clinical outcome of LAM patients by preventing loss of lung function. Nevertheless, other cell targets may be of interest for drug development. Therefore, we explored the potential role of EDN1 (endothelin) in LAM. We report an increased endothelin blood level in LAM patients as well as EDN1 overexpression and EDN1 receptor downregulation in LAM-derived primary cells and in TSC2

Topics: beta-Arrestin 1; Cell Line, Tumor; Cell Movement; Cell Proliferation; Endothelin A Receptor Antagonists; Endothelin-1; Female; Gene Expression Regulation, Neoplastic; Humans; Lung; Lung Neoplasms; Lymphangioleiomyomatosis; Primary Cell Culture; Receptor, Endothelin A; Receptor, Endothelin B; Signal Transduction; Sirolimus; Spheroids, Cellular; Tuberous Sclerosis; Tuberous Sclerosis Complex 2 Protein

Pulmonary metastases are the major cause of death of osteosarcoma (OS) patients. Endothelin-1 (ET-1) reportedly plays an important role in OS metastasis. In the present study, we for the first time explored the association of ET-1 SNPs with the risk of pulmonary metastatic OS. We genotyped three SNPs (rs1800541, rs2070699 and rs5370) in the ET-1 gene in a case-control study, using 260 pairs of age-, sex-, residence area- and tumor location-matched subjects. Patients with pulmonary metastatic OS and patients with localized high-grade (stage IIB) OS were enrolled as cases and controls, respectively. The G allele at rs1800541 was found associated with reduced risk of pulmonary metastatic OS after adjustment for body mass index, systolic blood pressure, diastolic blood pressure and the plasma ET-1 level (P=10(-4); adjusted OR, 0.55; 95% CI, 0.42-0.70), while the G allele at rs2070699 was not significantly associated with the risk of pulmonary metastatic OS (P=0.15; adjusted OR, 1.15; 95% CI, 0.87-1.50). The mRNA and the secreted protein levels of ET-1 in primary OS cell cultures (POCCs) established from surgically resected primary OS in the rs1800541 TT homozygotes were higher than those from the TG heterozygotes (P<0.05), who in turn showed higher ET-1 mRNA and secreted ET-1 levels than the GG homozygotes (P<0.05). In the control subjects, the rs1800541 TT homozygotes showed an 18.4% relapse rate, significantly higher than that of the GG homozygotes (0%) (P<0.01). On the other hand, the GG homozygotes showed a 71.4% complete recovery rate, significantly higher than that of the TG heterozygotes (7.3%) and the TT homozygotes (0%) (P<0.01). This study provides the first evidence of an association between the ET-1 gene SNPs and the risk of pulmonary metastatic OS.

Topics: Adolescent; Adult; Alleles; Case-Control Studies; Endothelin-1; Enzyme-Linked Immunosorbent Assay; Female; Gene Frequency; Genotype; Humans; Lung Neoplasms; Male; Osteosarcoma; Polymorphism, Single Nucleotide; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Young Adult

In the diagnosis of malignant mesothelioma (MM) there still is a lack of specific and sensitive screening biomarkers: this study examined the discriminatory power of a panel of serum/plasma biomarkers.. The study involved four groups: (a) individuals previously exposed to asbestos with asbestosis; (b) patients with MM; (c) patients with non-small cell lung cancer; and (d) controls without any evidence of malignancy. The concentrations of mesothelin, chitinase-3-like-1 (YKL-40), vascular endothelial growth factor (VEGF), endothelin-1, interleukin-8 (IL-8) and fibulin-3 in the serum of patients were determined.. Patients with MM had significantly higher serum levels of mesothelin (p<0.001), YKL-40 (p<0.001), IL-8 (p<0.001) and VEGF (p<0.01) than controls. The cut-off point for MM was 1.26 nM for mesothelin alone, and 167 pg/ml for YKL-40 alone; the presence of both markers above these cut-off levels improved diagnostic specificity.. The addition of YKL-40 may improve the specificity of mesothelin measurements alone for detecting patients with MM.

Topics: Adipokines; Aged; Asbestosis; Biomarkers, Tumor; Case-Control Studies; Chitinase-3-Like Protein 1; Cross-Sectional Studies; Endothelin-1; Female; GPI-Linked Proteins; Humans; Interleukin-8; Lectins; Lung Neoplasms; Male; Mesothelin; Mesothelioma; Middle Aged; Vascular Endothelial Growth Factor A

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

uPA and ET-1 proteins have been reported to be up-regulated in some of human cancers. The aim of this study is to investigate the alteration and clinical relevance of uPA and ET-1 protein levels in non-small cell lung cancer (NSCLC).. Expressions of uPA and ET-1 protein were detected in 155 cases of NSCLC with tissue microarrays and immunohistochemistry (TMA-IHC) technique. The correlations between the alteration of the two proteins and clinicopathological parameters were analyzed.. Negative/weak, moderate and high expression of uPA were observed in 12.3%, 64.4% and 23.3% of squamous cell carcinomas, in 12.2%, 53.7% and 34.1% of adenocarcinomas, and in 12.3%, 58.7% and 29.0% of all cases. ET-1 presented negative/weak, moderate and high expression in 2.7%, 42.5% and 54.8% of squamous cell carcinomas, in 11.0%, 30.5% and 58.5% of adenocarcinomas, and in 7.1%, 36.1% and 56.8% of all cases. Simultaneously high expression of uPA and ET-1 were found in adenocarcinomas without lymph node metastasis (P=0.017). Adenocarcinoma patients with high expression of uPA or with high expression of both ET-1 and uPA had the longer survival time (P=0.007 and 0.016).. Detection of uPA and ET-1 protein levels might contribute to the prognosis evaluation of NSCLC.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Carcinoma, Non-Small-Cell Lung; Endothelin-1; Female; Gene Expression Regulation, Neoplastic; Humans; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Up-Regulation; Urokinase-Type Plasminogen Activator

Two chemoprevention trials have shown that retinoic acid (RA) may be harmful in patients at risk for lung cancer, and RA administration to this high-risk group results in RARB2 reactivation. Although RARB2 is thought to possess tumor suppressive activity, its expression has recently been correlated with poorer prognosis in patients with nonsmall cell lung cancer. We hypothesized that RARB2 expression is necessary for the growth and maintenance of the oncogenic phenotype in lung cancer cells in which RARB2 has not been inactivated. We tested various antisense oligodeoxynucleotides (ASO) against RARB2 in multiple lung cancer cell lines and used microarray technology to compare the patterns of gene expression following ASO treatment versus RA treatment in the A-549 lung cancer cell line. We show that ASO treatment reduces proliferation and causes apoptosis in 3 RARB2-expressing lung cancer cell lines but has no apparent effect in at least two other lung cancer cells lines having lost RARB2 expression or one normal lung RARB2-expressing cell line; we demonstrate a correlation between resulting RARB2 expression levels and cell growth; and identify transcriptional effects related to both RA and RARB2 signaling. In particular, five genes known to contribute to carcinogenesis or chemotherapeutic resistance are down-regulated following ASO treatment: three of these are up-regulated following RA treatment. This work demonstrates a dual role for RARB2 (tumor suppression and tumor promotion) and identifies a challenge with respect to using RARB2 as a target for treatment or prevention strategies.

Topics: Antineoplastic Agents; Apoptosis; Cell Adhesion; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cyclooxygenase 2; Endothelin-1; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Humans; Interleukin-1beta; Lung Neoplasms; Receptors, Retinoic Acid; RNA Interference; Transcription, Genetic; Tretinoin

Regular use of beta(2)-adrenoceptor agonists may enhance non-specific airway responsiveness and inflammation. In earlier experimental studies, we showed that prolonged in vitro fenoterol exposure induced airway sensitization via perturbed epithelial regulation of bronchoconstriction. The aim of the present work was to examine the involvement of inflammatory mediator genes and proinflammatory cells and to investigate the role of the bronchial epithelium in these untoward effects. Bronchial tissues were surgically removed from 17 ex-smokers. Bronchial rings and primary cultures of bronchial epithelial cells were incubated with 0.1microM fenoterol for 15h. Levels of mRNA-expression were analyzed using a real-time quantitative reverse transcription-polymerase chain reaction array. Bronchial rings were contracted with endothelin-1 and immune cell infiltration was assessed by immunohistochemistry. Compared to paired controls, fenoterol up-regulated the mRNAs of cytokines/proteins implicated in the recruitment of T and B cells or the activation and proliferation of bronchial epithelial cells (CCL20/MIP-3alpha, FOXA2, PPAR-gamma) in isolated bronchi and in cultured epithelial cells. Fenoterol exposure significantly enhanced CD8(+)-T and differentiated CD138(+)-B-cells infiltration into the bronchi, especially the subepithelial area. Increase in CD8 or CD138 labeling-intensity strongly correlated with rise in maximal contraction to endothelin-1 induced by fenoterol exposure. In summary, our results show that fenoterol modulates the T and B cells chemotaxis possibly via the epithelial chemokine secretion in isolated bronchi from ex-smokers. They also suggest that the infiltration of resident T and B cells into the subepithelial area is associated with an increase in airway responsiveness due to fenoterol exposure.

Topics: Adrenergic beta-Agonists; Aged; B-Lymphocytes; Bronchi; Bronchial Hyperreactivity; Bronchoconstriction; Bronchoconstrictor Agents; CD8-Positive T-Lymphocytes; Cells, Cultured; Chemotaxis, Leukocyte; Cytokines; Dose-Response Relationship, Drug; Endothelin-1; Epithelial Cells; Female; Fenoterol; Gene Expression Regulation; Humans; Immunohistochemistry; Inflammation Mediators; Lung Neoplasms; Male; Middle Aged; Pulmonary Disease, Chronic Obstructive; Receptors, Adrenergic, beta; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Smoking Cessation; Time Factors

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

The DLX gene family encodes for homeobox transcription factors involved in the control of morphogenesis and tissue homeostasis. Their expression can be regulated by Endothelin1 (ET1), a peptide associated with breast cancer invasive phenotype. Deregulation of DLX gene expression was found in human solid tumors and hematologic malignancies. In particular, DLX4 overexpression represents a possible prognostic marker in ovarian cancer. We have investigated the role of DLX genes in human breast cancer progression.. MDA-MB-231 human breast carcinoma cells were grown in vitro or injected in nude mice, either subcutaneously, to mimic primary tumor growth, or intravenously, to mimic metastatic spreading. Expression of DLX2, DLX5 and DLX6 was assessed in cultured cells, either treated or not with ET1, tumors and metastases by RT-PCR. In situ hybridization was used to confirm DLX gene expression in primary tumors and in lung and bone metastases. The expression of DLX2 and DLX5 was evaluated in 408 primary human breast cancers examining the GSE1456 and GSE3494 microarray datasets. Kaplan-Meier estimates for disease-free survival were calculated for the patients grouped on the basis of DLX2/DLX5 expression.. Before injection, or after subcutaneous growth, MDA-MB-231 cells expressed DLX2 but neither DLX5 nor DLX6. Instead, in bone and lung metastases resulting from intravenous injection we detected expression of DLX5/6 but not of DLX2, suggesting that DLX5/6 are activated during metastasis formation, and that their expression is alternative to that of DLX2. The in vitro treatment of MDA-MB-231 cells with ET1, resulted in switch from DLX2 to DLX5 expression. By data mining in microarray datasets we found that expression of DLX2 occurred in 21.6% of patients, and was significantly correlated with prolonged disease-free survival and reduced incidence of relapse. Instead, DLX5 was expressed in a small subset of cases, 2.2% of total, displaying reduced disease-free survival and high incidence of relapse which was, however, non-significantly different from the other groups due to the small size of the DLX+ cohort. In all cases, we found mutually exclusive expression of DLX2 and DLX5.. Our studies indicate that DLX genes are involved in human breast cancer progression, and that DLX2 and DLX5 genes might serve as prognostic markers.

Topics: Animals; Biomarkers, Tumor; Bone Neoplasms; Breast Neoplasms; Cell Line, Tumor; Cell Movement; Disease-Free Survival; Endothelin-1; Female; Gene Expression Profiling; Homeodomain Proteins; Humans; In Situ Hybridization; Kaplan-Meier Estimate; Lung Neoplasms; Mice; Mice, Nude; Neoplasm Invasiveness; Oligonucleotide Array Sequence Analysis; Prognosis; Reverse Transcriptase Polymerase Chain Reaction; Time Factors; Transcription Factors

Angiogenesis is an essential process for solid tumor development. To interfere with angiogenesis, AdPPE3x-E1, an adenovirus that is transcriptionally targeted to replicate in angiogenic endothelial cells, was constructed, by replacing the E1 promoter with the modified preproendothelin-1 promoter, PPE-1-3x, previously shown to induce specific transcription in angiogenic endothelial cells.. The specificity of AdPPE3x-E1 to endothelial cells was shown by quantitative PCR and immunostaining, and its antiangiogenic effect was evaluated in Matrigel models. The in vivo efficacy of AdPPE3x-E1 was also tested in a cotton rat lung metastases model.. The replication rate of AdPPE3x-E1 in endothelial cells was similar to that of AdCMV-E1, a nonselective replicating adenovector, but the replication rate was reduced up to 60-fold in nonendothelial cells. Moreover, AdPPE3x-E1 reduced endothelial cell viability by 90% whereas nonendothelial cells were not affected. In in vitro and in vivo Matrigel models, endothelial cells infected with AdPPE3x-E1 did not develop capillary-like structures. The systemic administration of AdPPE3x-E1 reduced the lung metastases burden in a cotton rat model by 55%, compared with saline-treated rats, without significant evidence of toxicity. Quantitative PCR analysis showed that the viral copy number of AdPPE3x-E1 was increased 3-fold in the lung metastases but not in the liver, compared with a nonreplicating adenovector control.. We have shown here for the first time an antimetastatic effect induced by an angiogenesis-transcriptionally targeted adenovirus following systemic administration. Because adenovirus replication is more efficient in humans than in cotton rats, we assume a significant effect for AdPPE3x-E1 treatment in fighting human solid tumors and metastases.

Topics: Adenoviridae; Adenovirus E1 Proteins; Angiogenesis Inhibitors; Animals; Cells, Cultured; Collagen; DNA Replication; Drug Combinations; Endothelin-1; Endothelium, Vascular; Female; Fibroblasts; Humans; Kidney; Laminin; Lung Neoplasms; Neovascularization, Pathologic; Promoter Regions, Genetic; Proteoglycans; Rats; Sigmodontinae; Skin; Umbilical Veins; Virus Replication

Lung cancer is commonly associated with multiple-organ metastasis, and bone is a frequent metastatic site for lung cancer. Lung cancer frequently develops osteolytic, and less frequently osteoblastic, metastasis to bone. Osteolytic metastasis models of lung cancer have been reported, but no osteoblastic metastasis model is available for lung cancer. In the present study, we established a reproducible model of human lung cancer with both osteolytic and osteoblastic changes in natural killer cell-depleted severe combined immunodeficient mice. Intravenous inoculation of ACC-LC-319/bone2 cells resulted in the development of metastatic colonies in the lung, liver, and bone of the mice. As assessed sequentially by X-ray photographs, osteolytic bone lesions were observed by day 28, and then osteoblastic lesions were detected by day 35. Histological examination revealed the presence of bony spurs, a hallmark of osteoblastic bone metastasis, where osteoclasts were hardly observed. Treatment with an anti-human vascular endothelial growth factor antibody, bevacizumab, as well as zoledronate, inhibited the number of experimental bone metastases, including osteoblastic changes produced by ACC-LC-319/bone2 cells. These results indicate that our bone metastasis model by ACC-LC319/bone2 might be useful to understand the molecular pathogenesis of osteolytic and osteoblastic metastasis, and to identify molecular targets to control bone metastasis of lung cancer.

Topics: Adenocarcinoma; Animals; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Bevacizumab; Bone Neoplasms; Cell Line, Tumor; Diphosphonates; Disease Models, Animal; Endothelin-1; Humans; Imidazoles; Killer Cells, Natural; Lung Neoplasms; Male; Mice; Mice, Inbred ICR; Mice, SCID; Osteoblasts; Osteolysis; Vascular Endothelial Growth Factor A; Zoledronic Acid

Endothelin-1 (ET-1), the most potent vasoconstrictor, has been shown to be mitogenic in many tumor cells as well as in vascular cells. It was previously reported that the mRNA of ET-1 and endothelin receptors (ETRs) are expressed in lung cancer cells. However, their biological role in lung cancer remains to be explored. The purpose of this study was to determine whether ET-1 stimulates proliferation of the human lung adenocarcinoma cell SPC-A1 and probe its cellular mechanism. Reverse-transcription polymerase chain reaction and Western blot analysis showed that both the mRNA and protein of ET-1, ET A R and ET B R are expressed in SPC-A1 cells. Application of ET-1 at 10(-15)-10(-8) M caused a dose-dependent cell proliferation and an increase in intracellular free Ca2+ concentration ([Ca2+]i). This ET-1-induced cell proliferation and [Ca2+]i increase were completely abolished by BQ123, a selective ET A R antagonist, but not by BQ788, a selective ET B R antagonist. Furthermore, it was significantly reduced by U73122, a specific inhibitor of phospholipase C (PLC), but not by U73433, the structural isomer of U73122. Chelating extracellular Ca2+ or blocking voltage dependent calcium channels by nifedipine also significantly reduced the mitogenic effect of ET-1 and [Ca2+]i increase in SPC-A1 cells. These results indicate that ET-1 acts as an autocrine growth factor and enhances proliferation of SPC-A1 cells via activation of ET A R. The phosphoinositol/Ca2+ pathway and Ca2+ influx through voltage dependent Ca2+ channels activated by ET A R contribute to this process.

Topics: Blotting, Western; Calcium; Cell Line, Tumor; Cell Proliferation; Egtazic Acid; Endothelin-1; Humans; Lung Neoplasms; Nifedipine; Receptors, Endothelin; Reverse Transcriptase Polymerase Chain Reaction

Cytokines mediate numerous physiological and immune reactions, which are manifested in various biological effects, including tumouricidal activity. We evaluated the expression of the pleiotropic cytokine, tumour necrosis factor-alpha (TNF-alpha), by competitive PCR technique in 47 non-small cell lung cancer (NSCLC) cases and the impact of TNF-alpha on their clinical behaviour. Using univariate analysis, our study demonstrated a positive correlation between high TNF-alpha expression and favourable prognosis in NSCLC in terms of overall survival and disease free interval (p=0.03 and 0.04, respectively) and TNF-alpha maintained its independent role in multivariate analysis. TNF-alpha can stimulate the expression of many molecules, including interleukin-8 (IL-8) and endothelin-1 (ET-1); in our study, the expression of TNF-alpha was significantly associated with high IL-8 mRNA levels (p=0.008) and ET-1 mRNA positivity (p=0.03). We suggested that TNF-alpha can induce ET-1 mRNA expression in NSCLC, similarly to IL-8 expression. Our study may also contribute to advancing the knowledge of the molecular relationship between cytokines and endothelial functions in NSCLC.

Topics: Adult; Aged; Carcinoma, Non-Small-Cell Lung; Endothelin-1; Female; Gene Expression Regulation, Neoplastic; Humans; Interleukin-8; Lung Neoplasms; Male; Middle Aged; Multivariate Analysis; Prognosis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Survival Analysis; Tumor Necrosis Factor-alpha

The endothelin A receptor (ET(A)R) autocrine pathway is overexpressed in many malignancies, including nasopharyngeal carcinoma (NPC). In this tumor, ET(A)R expression is an independent determinant of survival and a robust independent predictor of distant metastasis. To evaluate whether ET(A)R represents a new target in NPC treatment, we tested the therapeutic role of ET(A)R in NPC. Cell proliferation was inhibited by the ET(A)R-selective antagonist ABT-627 in two ET(A)R-positive NPC cells in a dose-dependent manner. Proliferation of ET(A)R-negative NPC cells was not decreased. ET(A)R blockade also resulted in sensitization to cisplatin and 5-fluorouracil-induced apoptosis. In nude mice, ABT-627 inhibited the growth of NPC cell xenografts. Combined treatment of ABT-627 with the cytotoxic drug cisplatin or 5-fluorouracil produced additive antitumor effects. The antitumor activity of ABT-627 was demonstrated finally on an experimental lung metastasis by a reduction in the number of tumors. These results support the rationale of combining ABT-627 with current standard chemotherapy to further improve the therapeutic ratio in the treatment of NPC.

Topics: Animals; Antimetabolites, Antineoplastic; Antineoplastic Agents; Apoptosis; Atrasentan; Cell Proliferation; Cisplatin; Disease Models, Animal; Drug Therapy, Combination; Endothelin A Receptor Antagonists; Endothelin B Receptor Antagonists; Endothelin-1; Fluorouracil; Humans; Lung Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Nasopharyngeal Neoplasms; Pyrrolidines; Receptor, Endothelin A; Receptor, Endothelin B; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Neoplasm; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

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

The endothelin (ET) system influences tumourigenesis and tumour progression by various mechanisms, including angiogenesis. The aim of this study was to determine whether the expression of endothelin-1 (ET-1) is related to the angiogenic phenomenon in lung cancer and whether it could be involved in its clinical behaviour. Expression of ET-1, endothelin-converting enzyme-1 (ECE-1) and endothelin-receptors ETA and ETB was examined in 201 non-small cell lung carcinoma (NSCLC) and corresponding normal tissues using real-time polymerase chain reaction (RT-PCR). Forty NSCLC were also analysed for vascular endothelial growth factor (VEGF) expression by a competitive-PCR approach to assess whether ET-1 expression was related to this angiogenic factor. A higher number of cases with ET-1, ECE-1 and ETA mRNA expression was observed in malignant lung tumours compared with normal lung tissues (45.7% versus 33% for ET-1 (P < 0.0001); 38.3% versus 16.5% for ECE-1 (P = 0.004); and 42.8% versus 28.5% for ETA (P < 0.0001)). On the other hand, ETB mRNA was higher in normal lung tissues than in tumour samples (58.5% versus 52.8% (P < 0.0001)). Immunohistochemical analysis was also performed in 78 cases, selected from among those with high ET-1 mRNA, to confirm the presence of ET-1 protein and to determine its distribution and localisation. Moreover, an interesting relationship was observed between ET-1 and VEGF mRNA levels (P = 0.02). At univariate analysis, clinical-pathological parameters, such as sex, nodal metastatic involvement and stage, and ET-1 expression were seen to be significant predictors of worse prognosis regarding both overall survival (P = 0.001, P = 0.0003, P = 0.001 and P = 0.03, respectively) and disease-free interval (P = 0.0005, P = 0.0007, P = 0.001 and P = 0.04, respectively). We conclude that ET-1 could be involved in angiogenic phenomena in NSCLC and may represent a further indicator of progression and poor prognosis in this type of cancer, with interesting therapeutic implications.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Non-Small-Cell Lung; Disease-Free Survival; Endothelin-1; Female; Humans; Immunohistochemistry; Lung Neoplasms; Male; Middle Aged; Multivariate Analysis; Prognosis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Vascular Endothelial Growth Factor A

One recent line of cancer research is currently directed to the study of growth factors. Of increasing interest is endothelin-1 (ET-1), a mitogenic factor already investigated in several human cancer cell lines, which has been found to participate in the development and progression of tumours. This peptide has an important role also in non-small-cell lung cancer (NSCLC) where ET-1 expression has been found in 100% of cell lines.. The aim of this study was to measure ET-1 concentrations in the airways of patients with NSCLC using a completely non-invasive procedure--the breath condensate--and to verify the involvement of this peptide in the growth of lung tumours.. We enrolled 30 patients (17 men, median age 63 years; range 53-74) with histological evidence of NSCLC and 15 healthy controls (9 men, median age 59 years; range 52-70). ET-1 was measured in the exhaled breath condensate by means of a specific enzyme immunoassay kit.. Higher concentrations of exhaled ET-1 were found in NSCLC patients (8.3 +/- 0.7 pg/ml) compared to controls (5.2 +/- 0.5 pg/ml, p < 0.0001). A statistically significant difference was observed between patients with distant metastases (stage IV) of NSCLC (8.9 +/- 0.6 pg/ml) and those with locoregional disease (stage I-III) (7.9 +/- 0.5 pg/ml). A significant reduction in ET-1 levels was found in 14 patients after surgical removal of the tumour either associated with or without adjuvant chemotherapy (6.3 +/- 0.5 vs. 7.9 +/- 0.4 pg/ml, p < 0.0001).. These findings suggest that the measurement of ET-1 in the breath condensate of patients with NSCLC could be proposed as a marker for early detection of NSCLC as well as for monitoring reduction or progression of the neoplasm in the follow-up of treated patients.

Topics: Aged; Biomarkers, Tumor; Breath Tests; Carcinoma, Non-Small-Cell Lung; Early Diagnosis; Endothelin-1; Female; Humans; Lung Neoplasms; Male; Middle Aged; Predictive Value of Tests; Reproducibility of Results

Incubation of human distal bronchi from 48 patients for 15 h with 10(-7) M fenoterol induced sensitization characterized by an increase in maximal contraction to endothelin-1 (ET-1) and acetylcholine (ACh). Incubation of human bronchi with 10(-6), 3 x 10(-6), and 10(-5) M forskolin (an adenyl cyclase activator) reproduced sensitization to ET-1 and ACh. The sensitizing effect of fenoterol was inhibited by coincubation with gliotoxine (a nuclear factor-kappaB inhibitor), dexamethasone, indomethacin (a cyclooxygenase inhibitor), GR-32191 (a TP prostanoid receptor antagonist), MK-476 (a cysteinyl leukotriene type 1 receptor antagonist), SR-140333 + SR-48968 + SR-142801 (neurokinin types 1, 2, and 3 tachykinin receptor antagonists) with or without HOE-140 (a bradykinin B(2) receptor antagonist), SB-203580 (an inhibitor of the 38-kDa mitogen-activated protein kinase, p38(MAPK)), or calphostin C (a protein kinase C blocker). Our results suggest that chronic exposure to fenoterol induces proinflammatory effects mediated by nuclear factor-kappaB and pathways involving leukotrienes, prostanoids, bradykinin, tachykinins, protein kinase C, and p38(MAPK), leading to the regulation of smooth muscle contraction to ET-1 and ACh.

Topics: Acetylcholine; Adrenergic beta-Agonists; Albuterol; Bronchi; Colforsin; Endothelin-1; Ethanolamines; Female; Fenoterol; Formoterol Fumarate; Humans; In Vitro Techniques; Lung Neoplasms; Male; Middle Aged; Receptors, Adrenergic, beta-2

Gene therapy directed specifically to the vascular wall, particularly to angiogenic endothelial cells is a prerequisite in vascular disease treatment. Angiogenesis is a major feature in many pathological conditions including wound healing, solid tumors, developing metastases, ischemic heart diseases and diabetic retinopathy. In the present study we developed a tissue-specific gene therapy to the angiogenic blood vessels of tumor metastasis using an adeno-based vector containing the murine preproendothelin-1 (PPE-1) promoter. Genes activated by the PPE-1 promoter were highly expressed in bovine aortic endothelial cells in vitro. Systemic injection of the adenoviral vectors AdPPE-1-luciferase and AdCMV-luciferase to normal C57BL/6 mice, resulted in higher activity of PPE-1 promoter compared with CMV promoter in the aorta and vascularized tissues such as heart, kidney, lung and pancreas. Systemic administration of the adenoviral vector, in mice bearing Lewis lung carcinoma, resulted in high and specific activity of PPE-1 in the new vasculature of primary tumors and lung metastasis. Cellular distribution of the delivered gene revealed highest expression of GFP in angiogenic endothelial cells of the metastasis. We expect that this approach of 'vascular-directed' gene therapy will be applicable to both vascular diseases and cancer.

Topics: Adenoviridae; Analysis of Variance; Animals; Aorta; Carcinoma, Lewis Lung; Cattle; Cells, Cultured; Endothelin-1; Endothelins; Endothelium, Vascular; Gene Expression; Gene Targeting; Genetic Therapy; Genetic Vectors; Green Fluorescent Proteins; Liver; Luminescent Proteins; Lung Neoplasms; Male; Mice; Mice, Inbred C57BL; Microscopy, Fluorescence; Microscopy, Phase-Contrast; Neovascularization, Pathologic; Promoter Regions, Genetic; Protein Precursors; Statistics, Nonparametric

Cigarette smoking has been associated with alterations in the structure and endothelial function of pulmonary arteries. Nitric oxide (NO) and endothelin-1 are endothelium-derived mediators with opposite effects on vascular tone and cell growth. To investigate whether cigarette smoking could induce changes in the synthesis of these mediators in pulmonary arteries, we compared the expression of both endothelial NO synthase (eNOS) and endothelin-1 in the lungs of smokers with that in nonsmokers. Lung tissue samples of 23 smokers and nine nonsmokers were studied. Expression of eNOS and endothelin-1 in pulmonary artery endothelium was evaluated by immunohistochemistry. In protein extracts of lung tissue, the content of eNOS protein was assessed by Western blot analysis and that of endothelin-1 by radioimmunoassay. The immunohistochemical expression of eNOS in arterial endothelium and the eNOS protein content in lung tissue were lower in the smokers than in the nonsmokers. No differences were shown in cell expression and protein content of endothelin-1 between both groups. We conclude that cigarette smoking is associated with reduced expression of eNOS in pulmonary arteries. The diminished synthesis of nitric oxide may contribute to the alterations in the structure and endothelial function of pulmonary vessels in cigarette-smoke-induced respiratory disease.

Topics: Aged; Blotting, Western; Case-Control Studies; Endothelin-1; Endothelium, Vascular; Female; Gene Expression Regulation; Humans; Immunohistochemistry; Lung Neoplasms; Male; Middle Aged; Nitric Oxide Synthase; Pneumonectomy; Pulmonary Artery; Radioimmunoassay; Smoking

Aberrantly hypermethylated genes in human lung cancers were searched for by a genome scanning technique, methylation-sensitive-representational difference analysis (MS-RDA). A total of 59 DNA fragments were isolated as those methylated more heavily in either/both of two lung squamous cell carcinoma cell lines, EBC-1 and LK-2, than in a primary culture of normal human bronchial epithelium, NHBE. Thirty-four DNA fragments, whose hypermethylation was confirmed in primary squamous cell carcinomas, were sequenced. By database searches, 17 of them were shown to be located within 2 kb of putative CpG islands, and five of the 17 DNA fragments had transcribed regions of known genes in their vicinities. By RT-PCR of the five genes in the carcinoma cell lines and NHBE, decreased expression of HTR1B (5-hydroxytryptamine receptor 1B) and EDN1 (endothelin-1) was observed. Sequencing after bisulfite modification showed that the CpG island in the promoter region of HTR1B was hypermethylated, while that of EDN1 was not. Demethylation and re-expression of HTR1B were observed after treatment of LK-2 cells with 5-aza-2'-deoxycytidine. In primary lung cancers, decreased mRNA expression of HTR1B was observed in 11 of 20 cases, and that of EDN1 was in 16 of 20 cases. Immunohistochemical analysis of endothelin-1 confirmed that its immunoreactivity was reduced in squamous cell carcinoma cells compared with that in normal bronchial epithelial cells. Considering that endothelin-1 induces apoptosis in melanoma cells and that silencing of endothelin receptor B is observed in prostate cancers, its reduced expression was speculated to confer a growth advantage to lung cancer cells. MS-RDA was shown to isolate DNA fragments that are hypermethylated and silenced, such as HTR1B, and those whose expressions are altered and the methylation statuses outside the promoter region are altered, such as EDN1.

Topics: Aged; Blotting, Southern; Bronchi; Carcinoma, Squamous Cell; Cell Line; Cells, Cultured; CpG Islands; DNA; DNA Methylation; Endothelin-1; Epithelial Cells; Female; Gene Silencing; Humans; Immunohistochemistry; Introns; Lung Neoplasms; Male; Middle Aged; Models, Genetic; Physical Chromosome Mapping; Promoter Regions, Genetic; Receptor, Serotonin, 5-HT1B; Receptors, Serotonin; Reverse Transcriptase Polymerase Chain Reaction; Sequence Analysis, DNA; Sulfites; Transcription, Genetic; Tumor Cells, Cultured

Lung cancer, particularly small cell lung cancer (SCLC), is characterized by production of numerous peptides and their resulting clinical syndromes. Such peptides can act as autocrine growth factors for these tumors. In this study, we investigated the role of endothelin (ET)-1 in lung cancer. Using reverse transcription/polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay, and immunocytochemistry, we screened a panel of lung cancer cell lines for ET-1, its receptors, and endothelin converting enzyme-1 (ECE-1), which generates the active form of ET-1. ET-1 messenger RNA was expressed in five of seven SCLC, four of four non-small cell lung cancer (NSCLC), and human bronchial epithelial (HBE) cells. The intracellular isoform of ECE-1, important in processing ET-1 if an autocrine growth loop is to function, was downregulated in the lung cancer cell lines as compared with expression of the extracellular isoform. Endothelin A receptor (ETAR), which mediates the mitogenic effects of ET-1 in prostate and ovarian cancer, was upregulated in HBE cells compared with expression in three of seven SCLC and two of four NSCLC cell lines. Endothelin B receptor (ETBR) was more widespread, being expressed in seven of seven SCLC, four of four NSCLC, and the HBE cells. We used flow cytometry to measure mobilization of intracellular calcium as a functional assay for the ETAR. These data concurred with the RT-PCR results, indicating that the ETAR was downregulated or was involved in an alternative signal transduction pathway in lung cancer, and no evidence of functional receptor mediating an autocrine growth loop was found. From our study, the data do not support the putative functional autocrine growth role of ET-1 in lung cancer. We propose instead that ET-1 may act as a paracrine growth factor for surrounding epithelial and endothelial cells via alternative pathways, promoting angiogenesis and stromal growth.

Topics: Aspartic Acid Endopeptidases; Autocrine Communication; Bronchi; Calcium Signaling; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Endothelin-1; Endothelin-Converting Enzymes; Enzyme Induction; Enzyme-Linked Immunosorbent Assay; Epithelium; Flow Cytometry; Gene Expression Regulation, Neoplastic; Humans; Isoenzymes; Lung Neoplasms; Metalloendopeptidases; Microscopy, Fluorescence; Models, Biological; Neoplasm Proteins; Receptor, Endothelin A; Receptor, Endothelin B; Receptors, Endothelin; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Neoplasm; Tumor Cells, Cultured

The purpose of this study was to investigate a new method of in vivo gene transfer to the lung parenchyma by the percutaneous approach. The plasmid that contains the gene for firefly luciferase driven by a cytomegalovirus (CMV) promoter (pCMVL) in combination with cationic lipids was percutaneously injected into the lung parenchyma. Luciferase activities were localized to the lobes of the lung where the plasmids with cationic lipids were injected. Percutaneous injection of the plasmid containing the human endothelin-1 (hET-1) gene driven by a CMV promoter (pRc/CMVhET-1) in combination with cationic lipids into the lungs caused pulmonary fibrosis localized to the injection site in the peripheral lungs. We concluded that percutaneous in vivo gene transfer to the lungs is a unique and important approach to introduce exogenous gene expression in the limited area of the lung parenchyma. This method of gene transfer will be applicable for human gene therapy for targeted areas of peripheral lung and will also be useful to assess the function of the proteins expressed by a gene in the local area of the lungs.

Topics: Adenocarcinoma; Animals; Cytomegalovirus; Drug Carriers; Endothelin-1; Genes, Reporter; Humans; Liposomes; Luciferases; Lung; Lung Neoplasms; Male; Plasmids; Rats; Rats, Sprague-Dawley; Transfection; Tumor Cells, Cultured

This study was designed to characterize the endothelin pathway in an immortalized human adenocarcinoma-derived alveolar epithelial cell line (A549) and human umbilical vein endothelial cell line (HUVEC).. The release of ET-1 and big-ET-1 was measured in the incubation medium of both cell lines. The expression of mRNAs coding for the endothelin isoforms (hppET-1, -2, -3), the endothelin converting enzymes (hECE-1a, b, c, and d) and the hETA and hETB receptors was investigated using RT-PCR. The expression of ECE-1 mRNA in various human tissues and in A549 cells was investigated by Northern blot analysis and the subcellular localization of ECE-1 in A549 cells was investigated by immunoblotting using a polyclonal antibody.. Under control conditions, HUVEC release both ET-1 and big-ET-1 (ratio 5 to 1) while in A549 cells the big-ET-1 levels were below the threshold of detection. The release of these two peptides was minimally affected by various inhibitors of peptidases. However, in both cell lines phosphoramidon produced a concentration-dependent inhibition of ET-1 release and an enhanced accumulation of big-ET-1. Both HUVEC and A549 cells express the mRNAs for ppET-1, ET-A, and ET-B receptor subtypes and ECE-1 (isoforms ECE-1b, c and/or d). In addition, in HUVEC the mRNAs for ppET-2 and for the isoform ECE-1a were also detected. In A549 cells, ECE-1 had a preferential subcellular localization in the membrane fraction but was not detected in the cytosol.. Both A549 and HUVEC produce and release endothelin-1 through a specific enzymatic pathway, whether or not ECE-1 is the only enzyme involved remains to be determined. A549 might be used as a screening assay for drug discovery such as for inhibitors of endothelin-1 release.

Topics: Adenocarcinoma; Aspartic Acid Endopeptidases; Cells, Cultured; Endothelin-1; Endothelin-Converting Enzymes; Endothelins; Endothelium, Vascular; Glycopeptides; Humans; Lung Neoplasms; Metalloendopeptidases; Protein Precursors; RNA, Messenger; Tumor Cells, Cultured; Umbilical Veins

In order to study endothelin-1 (ET-1) positive expression in lung cancer and the relationship between the ET-1 quantitative analysis and the types, grades of lung cancer.. ET-1 positive expression and quantitative analysis were detected using avidin-biotin-peroxidase complex method and image analysis technology.. The ET-1 positive expression were mainly located in the cytoplasm in 104 cases with various types of lung cancer. The positive rate of ET-1 in adenocarcinoma, squamous-cell carcinoma and large-cell-lung cancer were 71.4%, 57.1% and 40.0% respectively. The positive rate of ET-1 in small-cell-lung cancer was 21.4%, significantly lower than others. The results of image analysis on adenocarcinoma and squamous-cell carcinoma showed that the lower lung cancer differentiation, the higher ET-1 quantitative.. There is ET-1 positive expression in all of types of lung cancer, but there is a high ET-1 positive expression in adenocarcinoma and squamous-cell carcinoma. The results of image analysis indicated that ET-1 quantitive expression was somehow related to the differentiation degree of neoplasm, so ET-1 quantitative analysis may be as a monitoring index of adenocarcinoma and squamous-cell carcinoma growing.

Topics: Adenocarcinoma; Carcinoma, Large Cell; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cell Differentiation; Endothelin-1; Humans; Lung Neoplasms

The mechanisms by which acute alveolar hypoxia induces pulmonary vasoconstriction remain unclear. We investigated whether endothelin-1 (ET-1) could be detected in plasma during pulmonary alveolar hypoxia without systemic hypoxemia (one-lung hypoxia) and whether the levels could be related to hemodynamic status in humans.. Thirteen adult patients with primary lung carcinoma were studied prior to surgery. Anesthesia was induced with fentanyl, diazepam, and pancuronium iv. Differential lung ventilation was performed for 40 min. The right lung was ventilated with a mixture of 6% O2, 5% CO2, and 89% N2 and the left lung ventilated with 100% O2. Blood gas values, hemodynamic parameters, and plasma ET-1 levels were measured.. Mean pulmonary artery pressure increased from 13.0 +/- 3.5 to 17.2 +/- 3.2 mm Hg (P < 0.01) after 20 min of one-lung hypoxia. The plasma ET-1 levels in arterial blood and wedged right pulmonary arterial blood increased from 1.69 +/- 0.61 to 2.13 +/- 0.48 pg/ml (P < 0.01) and from 1.75 +/- 0.47 to 2.26 +/- 0.40 pg/ml (P < 0.001), respectively, after 40 min of one-lung hypoxia. At baseline and after 40 min of one-lung hypoxia, there was a significant correlation between the mean pulmonary artery pressure and arterial plasma ET-1 levels (r = 0.650, P < 0.01) in the wedged pulmonary arterial plasma levels (r = 0.484, P < 0.05). The increase in plasma ET-1 levels in arterial blood and in the wedged pulmonary arterial blood of the hypoxic lung occurred slowly.. We concluded that ET-1 may a supporting, but not a primary, role in human hypoxic pulmonary vasoconstriction.

Topics: Aged; Anesthesia; Blood Pressure; Carbon Dioxide; Endothelin-1; Female; Humans; Hypoxia; Lung Neoplasms; Male; Middle Aged; Oxygen; Pneumonectomy; Pulmonary Alveoli; Pulmonary Artery; Respiration, Artificial; Vascular Resistance

1. We have studied the contractile activity of the 39 amino acid precursor of endothelin-1 (ET-1), big endothelin-1 (big ET-1), on human isolated bronchi. The contribution of the metalloproteases, neutral endopeptidase (NEP) and angiotensin converting enzyme (ACE), in the presence or absence of the epithelium lining, by use of specific inhibitors, was also evaluated on the effects of big ET-1. 2. Big ET-1 elicited a potent contraction of human isolated bronchus. The -log EC50 value for big ET-1 was 7.53 +/- 0.08 (n = 11) and Emax 78.5 +/- 3.8% (% of ACh 3mM). 3. Incubation of human isolated bronchi with the NEP inhibitor phosphoramidon (10(-5) M) induced a rightward shift of the concentration-response curve induced by big ET-1 (10(-9) M to 3 x 10(-7) M). Similar results were observed when human bronchi were incubated with thiorphan (10(-5) M), but the shift to the right was significantly less (P less than 0.01) than that observed in the case of phosphoramidon (-0.35 +/- 0.05 vs -0.67 +/- 0.07 log unit). 4. The two inhibitors of angiotensin I converting enzyme (ACE), captopril or enalapril diacid, did not affect the concentration-response curve for contraction induced by big ET-1. 5. When the epithelium was removed, a leftward shift of the concentration-response curve of big ET-1 (10(-9) M to 3 x 10(-7) M) was observed. Incubation of human isolated bronchi with phosphoramidon or thiorphan (10-5M) or with enalapril diacid or captopril did not modify the leftward shift of the concentration-response curve for big ET-1 after epithelium removal.6. These results suggest that big ET-1 elicits potent contractile activity in the human isolated bronchus and that its effect is the consequence of the conversion to ET-1 by a phosphoramidon-sensitive metalloprotease which, although different from NEP and ACE, appears to be similar to the endothelinconverting enzyme (ECE) described in other studies in animals.

Topics: Angiotensin-Converting Enzyme Inhibitors; Bronchi; Dose-Response Relationship, Drug; Endothelin-1; Endothelins; Endothelium; Female; Humans; In Vitro Techniques; Lung Neoplasms; Male; Middle Aged; Muscle Contraction; Neprilysin; Protein Precursors