elastin and Lung-Neoplasms

Elastin and collagen are the main components of the lung connective tissue network, and together provide the lung with elasticity and tensile strength. In pulmonary pathology, elastin staining is used to variable extents in different countries. These uses include evaluation of the pleura in staging, and the distinction of invasion from collapse of alveoli after surgery (iatrogenic collapse). In the latter, elastin staining is used to highlight distorted but pre-existing alveolar architecture from true invasion. In addition to variable levels of use and experience, the interpretation of elastin staining in some adenocarcinomas leads to interpretative differences between collapsed lepidic patterns and true papillary patterns. This review aims to summarise the existing data on the use of elastin staining in pulmonary pathology, on the basis of literature data and morphological characteristics. The effect of iatrogenic collapse and the interpretation of elastin staining in pulmonary adenocarcinomas is discussed in detail, especially for the distinction between lepidic patterns and papillary carcinoma.

Topics: Adenocarcinoma of Lung; Adenocarcinoma, Papillary; Collagen; Diagnosis, Differential; Elastin; Histocytochemistry; Humans; Lung Neoplasms; Pleura; Pulmonary Alveoli

Human neutrophil elastase (HNE), a main actor in the development of chronic obstructive pulmonary diseases, has been recently involved in non-small cell lung cancer progression. It can act at several levels (i) intracellularly, cleaving for instance the adaptor molecule insulin receptor substrate-1 (IRS-1) (ii) at the cell surface, hydrolyzing receptors as CD40 (iii) in the extracellular space, generating elastin fragments i.e. morphoelastokines which potently stimulate cancer cell invasiveness and angiogenesis. Since decades, researchers identified natural compounds and/or synthesized agents which antagonize HNE activity that will be described in this review article. Some of these compounds might be of value as therapeutic agents in lung cancer. However, it is now widely accepted that lung tumor invasion and metastasis involve proteolytic cascades. Accordingly, we will here mainly focus our attention to natural substances able to display a dual inhibitory capacity (i.e. lipids and derivatives, phenolics) towards HNE and matrix metalloproteinases (MMPs), particularly MMP-2. To that purpose, we recently synthesize substances named "LipoGalardin" (Moroy G. et al., Biochem. Pharmacol., 2011, 81(5), 626-635) exhibiting such inhibitory bifunctionality. At last, we will propose an original synthetic scheme for designing a potent biheaded HNE/MMP-2 inhibitor.

Topics: alpha 1-Antitrypsin; Antineoplastic Agents; Carcinoma, Non-Small-Cell Lung; CD40 Antigens; Elastin; Enzyme Inhibitors; Humans; Insulin Receptor Substrate Proteins; Leukocyte Elastase; Lung Neoplasms; Matrix Metalloproteinase Inhibitors; Matrix Metalloproteinases; Models, Molecular

This mini-review summarizes our recent experimental and clinical studies on neutrophil elastase (NE) and cancer based on our original view point. Neoplasms metastasize as a result of a complex series of events. This process requires various degradative enzymes including proteases. NE has broad substrate specificity under physiological conditions, and excessive NE results in digestion of not only elastin, but also other extracellular matrix proteins. Several cell lines from human breast cancer and human lung cancer produce immunoreactive NE. The amount of immunoreactive NE in tumor tissue is an independent prognostic indicator of patients with breast cancer and lung cancer. Furthermore, a specific NE inhibitor completely suppressed growth of cancer cells transplanted into severe combined immunodeficiency mice. The use of NE inhibitor would seem to be a promising way to prevent the invasion and metastasis of cancer.

Topics: Animals; Breast Neoplasms; Elastin; Enzyme Inhibitors; Humans; Leukocyte Elastase; Lung Neoplasms; Neoplasm Metastasis; Prognosis

It is now well established that the interaction of tumour cells with elastin is important during invasion and metastasis. This is due to the fact that the elastin receptor complex is widely expressed by tumour cells and is overexpressed in highly metastatic variants. There is evidence that the elastin receptor complex is associated with a signal system involving G proteins, phospholipase C, the phosphoinositol cycle and protein kinase C. Therefore, activation of the elastin receptor system results in activation of protein kinase C-dependent cellular processes such as enzyme secretion and migration. Accordingly, soluble elastin can be used in vivo to interfere with tumour cell dissemination into elastin-rich tissues such as lung, skin or blood vessels. The importance of elastin-tumour cell interactions is emphasized by the observation that the 67 kDa receptor for laminin may well be identical to the 67 kDa elastin receptor of the elastin receptor complex. Interference with the function of this receptor system by the use of both laminin peptides and elastin ligands may provide the basis for a novel and more powerful antimetastatic intervention.

Topics: Amino Acid Sequence; Animals; Elastin; Humans; Lung Neoplasms; Molecular Sequence Data; Neoplasm Metastasis; Neoplasms; Receptors, Cell Surface

Metastasis is a multistep process in which amoeboid chemotaxis plays a key role in the movement of tumor cells into and out of vessels. On a molecular level, much of what is known about amoeboid chemotaxis has been learned through work with Dictyostelium discoideum, a lower eukaryotic amoeboid phagocyte. One of the first and most crucial events to occur in the actin cytoskeleton following chemotactic stimulation is activation of actin nucleation. This is followed by incorporation of specific actin cross-linking proteins into the cytoskeleton, proteins which are implicated in the extension of pseudopods and filopods. Together, these events have been termed the Cortical Expansion Model for amoeboid chemotaxis. Detailed biochemical analysis has implicated a new actin-capping protein and has shown that one of the cross-linking proteins is Elongation Factor 1a, suggesting a link between chemotaxis and growth control. Preliminary data from parallel studies on neoplastic cells are presented.

Topics: Amino Acid Sequence; Animals; Chemotaxis; Dictyostelium; Elastin; Lung Neoplasms; Mice; Molecular Sequence Data; Neoplasm Metastasis; Oligopeptides; Phagocytes

In this review article, the effects of old age on lung structure and function are discussed. Changes in lung morphology and biochemistry are correlated with changes in lung mechanics and gas exchange, as well as with the respiratory system's adaptability to the stresses of exercise and sleep. The effects of aging on the lungs' defense mechanisms are related to pulmonary diseases of the elderly.

Topics: Adaptation, Physiological; Aging; Animals; Asthma; Carcinoma, Bronchogenic; Closing Volume; Collagen; Diaphragm; Elastin; Humans; Lung; Lung Neoplasms; Maximal Expiratory Flow-Volume Curves; Nucleic Acids; Pneumonia; Proteins; Proteoglycans; Pulmonary Circulation; Pulmonary Diffusing Capacity; Pulmonary Gas Exchange; Residual Volume; Respiration; Total Lung Capacity; Tuberculosis, Pulmonary; Vital Capacity

Elastin is a signature protein of lungs. Increased elastin turnover driven by altered proteolytic activity is an important part of lung tumorigenesis. Elastin-derived fragments have been shown to be pro-tumorigenic, however, little is known regarding the biomarker potential of such elastin fragments. Here, we present an elastin turnover profile by non-invasively quantifying five specific elastin degradation fragments generated by different proteases.. Elastin fragments were assessed in serum from patients with stage I-IV non-small cell lung cancer (NSCLC) (n = 40) and healthy controls (n = 30) using competitive ELISAs targeting different protease-generated fragments of elastin: ELM12 (generated by matrix metalloproteinase MMP-9 and -12), ELM7 (MMP-7), EL-NE (neutrophil elastase), EL-CG (cathepsin G) and ELP-3 (proteinase 3).. ELM12, ELM7, EL-NE and EL-CG were all significantly elevated in NSCLC patients (n = 40) when compared to healthy controls (n = 30) (ELM12, p = 0.0191; ELM7, p < 0.0001; EL-NE, p < 0.0001; EL-CG, p < 0.0001). ELP-3 showed no significant difference between patients and controls (p = 0.8735). All fragments correlated positively (Spearman, r: 0.69-0.81) when compared pairwise, except ELM12 (Spearman, r: 0.042-0.097). In general, all fragments were detectable across all stages of the disease.. Elastin fragments generated by different proteases are elevated in lung cancer patients compared to healthy controls but differ in their presence. This demonstrates non-invasive biomarker potential of elastin fragments in serum from lung cancer patients and suggests that different pathological mechanisms may be responsible for the elastin turnover, warranting further validation in clinical trials.

Topics: Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Case-Control Studies; Elastin; Female; Follow-Up Studies; Humans; Lung; Lung Neoplasms; Male; Middle Aged; Prognosis

Topics: A549 Cells; Administration, Inhalation; Carcinoma, Non-Small-Cell Lung; Cell Proliferation; Chitosan; Curcumin; Doxorubicin; Drug Carriers; Drug Liberation; Elastin; Humans; Hydrogen-Ion Concentration; Lung Neoplasms; Microspheres; Particle Size; Solubility

During the pathological destruction of lung tissue, neutrophil elastase (NE) degrades elastin, one of the major constituents of lung parenchyma. However there are no non-invasive methods to quantify NE degradation of elastin. We selected specific elastin fragments generated by NE for antibody generation and developed an ELISA assay (EL-NE) for the quantification of NE-degraded elastin.. Monoclonal antibodies were developed against 10 NE-specific cleavage sites on elastin. One EL-NE assay was tested for analyte stability, linearity and intra- and inter-assay variation. The NE specificity was demonstrated using elastin cleaved in vitro with matrix metalloproteinases (MMPs), cathepsin G (CatG), NE and intact elastin. Clinical relevance was assessed by measuring levels of NE-generated elastin fragments in serum of patients diagnosed with idiopathic pulmonary fibrosis (IPF, n = 10) or lung cancer (n = 40).. Analyte recovery of EL-NE for human serum was between 85% and 104%, the analyte was stable for four freeze/thaw cycles and after 24 h storage at 4°C. EL-NE was specific for NE-degraded elastin. Levels of NE-generated elastin fragments for elastin incubated in the presence of NE were 900% to 4700% higher than those seen with CatG or MMP incubation or in intact elastin. Serum levels of NE-generated elastin fragments were significantly increased in patients with IPF (137%, p = 0.002) and in patients with lung cancer (510%, p < 0.001) compared with age- and sex-matched controls.. The EL-NE assay was specific for NE-degraded elastin. The EL-NE assay was able to specifically quantify NE-degraded elastin in serum. Serum levels of NE-degraded elastin might be used to detect excessive lung tissue degradation in lung cancer and IPF.

Topics: Adenocarcinoma; Aged; Carcinoma, Squamous Cell; Case-Control Studies; Elastin; Enzyme-Linked Immunosorbent Assay; Extracellular Matrix; Female; Humans; Idiopathic Pulmonary Fibrosis; Leukocyte Elastase; Lung; Lung Neoplasms; Male; Middle Aged; Peptide Fragments; Small Cell Lung Carcinoma

Elastin is a signature protein of the lungs. Matrix metalloproteinase-7 (MMP-7) is important in lung defence mechanisms and degrades elastin. However, MMP-7 activity in regard to elastin degradation has never been quantified serologically in patients with lung diseases. An assay for the quantification of MMP-7 generated elastin fragments (ELM7) was therefore developed to investigate MMP-7 derived elastin degradation in pulmonary disorders such as idiopathic pulmonary fibrosis (IPF) and lung cancer.. Monoclonal antibodies (mABs) were raised against eight carefully selected MMP-7 cleavage sites on elastin. After characterisation and validation of the mABs, one mAB targeting the ELM7 fragment was chosen. ELM7 fragment levels were assessed in serum samples from patients diagnosed with IPF (n=123, baseline samples, CTgov reg. NCT00786201), and lung cancer (n=40) and compared with age- and sex-matched controls.. The ELM7 assay was specific towards in vitro MMP-7 degraded elastin and the ELM7 neoepitope but not towards other MMP-7 derived elastin fragments. Serum ELM7 levels were significantly increased in IPF (113%, p<0.0001) and lung cancer (96%, p<0.0001) compared to matched controls.. MMP-7-generated elastin fragments can be quantified in serum and may reflect pathological lung tissue turnover in several important lung diseases.

Topics: Aged; Animals; Case-Control Studies; Elastin; Female; Humans; Idiopathic Pulmonary Fibrosis; Lung; Lung Diseases; Lung Neoplasms; Male; Matrix Metalloproteinase 7; Mice; Mice, Inbred BALB C; Middle Aged; Proteolysis

Targeted therapy focused on highly expressed growth factor receptors is increasingly becoming popular for the treatment of lung cancer. Cancer cells exhibit higher levels of macropinocytosis than the normally quiescent non-cancerous cells, which can further be enhanced by growth factors. Here, we show the targeted enhancement of macropinocytosis in lung cancer cells for the delivery of the mitochondriotoxic peptide (KLAKLAK)2 using keratinocyte growth factor (KGF). We report the formation of a nanoparticle (NP) comprising of two chimeric fusion proteins, both fused to elastin-like polypeptide (ELP), (KLAKLAK)2-ELP and KGF-ELP. We show that (KLAKLAK)2-ELP nanoparticles are internalized via macropinocytosis and its internalization is facilitated by the interaction of the ELP domain with cell surface heparin sulfate proteoglycans. This internalization leads to mitochondrial depolarization and subsequent cell death. Also, we demonstrate that KGF-ELP selectively enhances macropinocytosis in cancer cells expressing high levels of the keratinocyte growth factor receptor (KGFR). Finally, the heterogeneous NPs consisting of (KLAKLAK)2-ELP and KGF-ELP selectively kill KGFR-expressing lung cancer cells. Hence, this multipronged approach of targeting highly active processes and receptors in cancer cells will be tremendously selective in the treatment of both early-stage and advanced-stage lung cancers, thereby improving patient's prognosis and survival rate.

Topics: Antineoplastic Agents; Cell Line, Tumor; Drug Delivery Systems; Elastin; Humans; Lung Neoplasms; Pinocytosis; Receptor, Fibroblast Growth Factor, Type 2; Recombinant Fusion Proteins

Surgical removal and pathologic handling of lung tissue has a compressive effect upon its architecture. The effect of surgical atelectasis on morphology has not been examined in depth, especially with respect to lung adenocarcinomas.. To examine the influence of surgical atelectasis on morphologic lepidic growth pattern, mimicking papillary adenocarcinoma pattern.. In 2 cases serial sections of resected pulmonary adenocarcinoma were used, as was a 3-dimensional reconstruction. Elastin stains were performed on primary and metastatic adenocarcinomas.. Perfusion fixation of another case showed marked morphologic differences of less compressed peripheral lung tissue, emphasizing the preexisting alveolar structure. An elastic stain may help identify true lesional architecture.. We demonstrate that microscopic sections of adenocarcinoma in situ in compressed/collapsed tissue may give rise to a pseudopapillary pattern mimicking invasive adenocarcinoma. Accurate appreciation of different tumor architecture in lung adenocarcinoma has important biologic and clinical implications. Pathologists should be aware of the possibility of misclassification of adenocarcinoma pattern due to tissue artifacts caused by lung tissue handling.

Topics: Adenocarcinoma; Adenocarcinoma, Papillary; Aged, 80 and over; Artifacts; Biomarkers, Tumor; Diagnosis, Differential; Elastin; Female; Histocytochemistry; Humans; Lung; Lung Neoplasms; Male; Middle Aged; Specimen Handling; Tomography, X-Ray Computed

Elastin-rich lung extracellular matrix is largely remodeled during tumor invasion. Elastin degradation produces peptides displaying a wide range of biological activities. These elastin derived peptides (EP) interact with the elastin receptor complex (ERC) but also bind to α(V)β(3) integrin and galectin-3. In this study, we explored the role of EP and their receptors in tumor progression of lung carcinomas. Non-invasive and invasive lung tumor cell lines were incubated in presence of kappa-elastin (κE) or with synthetic peptides displaying receptor-specific sequences (VGVAPG, GRKRK, AGVPGLGVG and AGVPGFGAG). Modified Boyden chamber assays revealed an increased invasive capacity of invasive cells induced by κE. EP treatment had no effect on cell proliferation but zymography analysis revealed an increase of pro-MMP-2 and uPA levels in the conditioned media of treated cells. Moreover, the active form of MMP-2 was increased in invasive cells. Interestingly, this regulation was not observed at the mRNA level and actinomycin D was unable to inhibit κE effects. We also observed that the regulation of proteases protein level following κE treatment was an early process detectable after 1 h. All these effects could not be inhibited by lactose and V14, two ERC antagonists, or by blocking antibodies against α(V)β(3) integrin and galectin-3. Finally, VGVAPG and GRKRK failed to reproduce κE effects whereas nonapeptides partially mimicked them. These results demonstrate that treatment with EP up-regulates invasiveness of lung tumor cells via the release of proteolytic enzymes. This modulation involves post-transcriptional mechanisms and a nonapeptide-receptor different from the ERC, α(V)β(3) integrin and galectin-3.

Topics: Adenocarcinoma; Biomarkers, Tumor; Blotting, Western; Cell Movement; Cell Proliferation; Elastin; Extracellular Matrix; Galectin 3; Gene Expression Profiling; Humans; Integrin alphaVbeta3; Lung Neoplasms; Matrix Metalloproteinase 2; Oligonucleotide Array Sequence Analysis; Oligopeptides; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA Processing, Post-Transcriptional; RNA, Messenger; Urokinase-Type Plasminogen Activator

Non-small cell lung carcinoma (NSCLC) is a highly fibrotic malignancy, which exhibits a prominent desmoplastic stroma. Epithelial-mesenchymal transition (EMT) is one of the main modes of carcinoma invasion. We identified the stromal N-glycoprotein periostin by mass spectrometry of lung adenocarcinoma pleural effusions. Validation on a NSCLC tissue microarray and on tumor whole sections by immunohistochemistry indicated that periostin is strongly upregulated at the invasive front in both tumor epithelia and the surrounding matricellular space. In comparison to collagen, elastin and vimentin, periostin was found to be most closely associated with parameters of tumor progression such as larger size and higher stage, with the squamous cell histotype, and with decreased survival. An association with decreased survival was also found for the cell adhesion molecule L1CAM. In conclusion, enlargement of NSCLC tumors is associated with an increase of desmoplastic stroma and concomitant upregulation of EMT markers at the invasive front. The tumor-stroma interface may be a candidate topographic region for stroma- or EMT-directed therapy.

Topics: Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Cell Adhesion Molecules; Collagen; Disease Progression; Disease-Free Survival; Elastin; Epithelial-Mesenchymal Transition; Humans; Lung; Lung Neoplasms; Mass Spectrometry; Neoplasm Invasiveness; Neural Cell Adhesion Molecule L1; Oligonucleotide Array Sequence Analysis; Up-Regulation; Vimentin

To investigate a unique biomarker from the blood plasma and sputum of lung cancer patients based on native fluorescence analysis of body fluids.. Conventionally, biomarkers indicative of malignancy are identified by biochemical or biophysical processes. Most of the cancer biomarkers, often useful in monitoring disease progression, have sensitivity and specificity in the range of 60%.. We employed synchronous fluorescence excitation spectroscopy (SFXS) and fluorescence emission spectroscopy for the native fluorescence analysis of blood plasma of 32 normal controls, 32 patients with lung cancer, and 32 patients with other types of cancer.. Based on the native fluorescence analysis of blood plasma and sputum, we found that the structural protein elastin, with an excitation peak at 327 nm and an emission peak at 405 nm, is an exclusive biomarker for lung carcinoma with 77% sensitivity and 83% specificity from plasma alone, 92.3% sensitivity and 100% specificity from plasma acetone extract alone, and 66% sensitivity and 100% specificity from sputum alone.. In this preliminary report with a limited number of lung cancer patients, we have used SFXS of plasma and sputa as the basis for a new technique identifying elastin as an exclusive lung cancer biomarker. This technique has the potential to become a new protocol for rapid and cost-effective screening and diagnosis of lung cancer.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Case-Control Studies; Elastin; Female; Humans; Lung Neoplasms; Male; Middle Aged; Reference Values; Sensitivity and Specificity; Spectrometry, Fluorescence; Sputum

Desmosine (DES) and isodesmosine (IDES) are both pyridinium amino acid isomers that serve as cross-linking molecules binding the polymeric chains of amino acids into elastin. Found in urine, they are markers for the degradation of elastin which occurs in chronic obstructive pulmonary disease (COPD). In this study, a robust method using ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) with selected reaction monitoring (SRM) mode was developed for the analysis of DES and IDES in human urine. Pyridylethyl-cysteine (PE-Cys) as internal standard (I.S.) was employed for the quantification of DES and IDES. The analytes and I.S. were extracted by solid-phase extraction with Oasis MCX cartridges and separated on an AccQ-Tag Ultra column. The assay was accurate (-6.8% to 14.5%) and precise (2.8% to 13.8%) within the concentration range of 1 to 250 pmol/mL. Moreover, the recovery and stability (working/ I.S. solution, urine samples with added elastin, and pretreated sample) was investigated, and these parameters were found acceptable. The UPLC-MS/MS method was validated and had good reproducibility and stability for the quantification of DES and IDES, which requires only 100 mL of human urine. This assay will be a useful means for measuring DES and IDES levels in urine with robustness and characterizing patients with COPD.

Topics: Adult; Calibration; Case-Control Studies; Chromatography, Liquid; Desmosine; Drug Stability; Efficiency; Elastin; Female; Humans; Isodesmosine; Lung Neoplasms; Lymphangioleiomyomatosis; Male; Models, Biological; Reproducibility of Results; Sensitivity and Specificity; Specimen Handling; Tandem Mass Spectrometry; Urinalysis

In carcinomas, invasive tumor growth is accompanied by desmoplastic stroma reaction and facilitated by epithelial-mesenchymal transition (EMT) of cancer cells. We investigated the prognostic significance of the EMT indicator proteins periostin and vimentin in comparison with versican, a putative indicator of the opposite mechanism mesenchymal-epithelial transition (MET), and to the desmoplasia proteins collagen and elastin in non-small cell lung cancer (NSCLC).. Tumor of 533 patients with surgically resected NSCLC was used for analysis of stromal and epithelial protein expression by immunohistochemistry (EMT-MET proteins) and Elastica van Gieson histochemical staining (collagen and elastin). A semiquantitative sum scoring system was done on three tissue microarrays.. Of the 533 patients, 48% had squamous cell carcinoma, 47% adenocarcinoma, and 5% adenosquamous carcinoma. High expression of periostin in either stroma or tumor epithelia, independently scored by two pathologists, correlated with male gender, higher stage, higher pT category, and larger tumor size, and in only stroma with tumor relapse. High expression of versican in either stroma or epithelia as well as of stromal collagen had fewer but concordant associations with advanced tumor and periostin, respectively. High expression of elastin was oppositely associated with less advanced disease. Associations of high vimentin were inconsistent (all P values < 0.05). High stromal periostin was found to be a prognostic factor for decreased progression-free survival on univariate analysis (P = 0.007).. Because up-regulation is frequently observed in the stromal and epithelial tumor compartment, EMT-MET indicator proteins may be integrated in progression models of NSCLC.

Topics: Age Factors; Aged; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Cell Adhesion Molecules; Cell Differentiation; Collagen; Elastin; Epithelial Cells; Female; Humans; Immunohistochemistry; Kaplan-Meier Estimate; Lung Neoplasms; Male; Mesoderm; Middle Aged; Prognosis; Sex Factors; Tissue Array Analysis; Versicans; Vimentin

The interstitial lung disease lymphangioleiomyomatosis (LAM) is characterized by diffuse proliferation of smooth muscle cells (SMCs), which in many patients show TSC2 (tuberin) gene mutations, in addition to thickening of interstitial tissues, loss of alveoli, and the development of cystic spaces. While SMC proliferation is the defining feature of LAM, a significant proportion of LAM lung tissue consists of expanded interstitial connective tissue that is negative for smooth muscle actin and TSC2 mutations. The importance of this actin-negative interstitial tissue to the pathophysiology of LAM is not clear. The present study has determined the contribution of this interstitial tissue to LAM lung volume by morphometric analysis and has examined its cell and matrix proteoglycan composition by immunohistochemistry. Lung tissue from nine LAM patients and four control subjects was examined. LAM lung contained twice as much interstitial tissue as control lung (27% versus 13% of total lung volume), with SMCs accounting for less than 25% of the interstitial volume. Areas of interstitial tissue stained strongly for the matrix proteoglycans versican and biglycan. Decorin was prominent in association with collagen bundles. SMCs did not stain, or stained lightly, for proteoglycans. Versican and biglycan deposits were closely associated with actin-negative interstitial fibroblasts identified by prolyl 4-hydroxylase immuno-staining. Comparatively normal alveolar walls in LAM lung also stained strongly for versican and had a reduced elastin content. Thickened interstitial regions contained significant amounts of elastin (approximately 13% of interstitial volume) but with fibres in disorganized patterns. Elastic fibres were absent from areas that stained strongly for versican and biglycan. These areas also showed weak staining for elastin binding protein (EBP), consistent with proteoglycan-induced shedding of EBP and inhibition of elastic fibre formation. These findings point to a significant contribution from matrix proteoglycans to the expanded and remodelled interstitial lung tissue of LAM patients.

Topics: Actins; Biglycan; Chondroitin Sulfate Proteoglycans; Collagen; Decorin; Elastin; Extracellular Matrix; Extracellular Matrix Proteins; Fibroblasts; Humans; Immunohistochemistry; Lectins, C-Type; Lung; Lung Neoplasms; Lymphangioleiomyomatosis; Muscle, Smooth; Proteoglycans; Pulmonary Alveoli; Receptors, Cell Surface; Versicans

Elastin metabolism parameters (elastin-derived peptides and elastase-like activity) were determined in sera of patients with lung cancer and in healthy controls. The concentration of elastin-derived peptides was statistically significantly elevated in the lung cancer group. There was no statistically significant difference in the serum elastase-like activity between the groups studied. These data seem to indicate an enhanced metabolism of elastin in patients with lung cancer.

Topics: Adenocarcinoma; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Elastin; Female; Humans; Lung Neoplasms; Male; Middle Aged; Pancreatic Elastase; Peptide Fragments

The aim of this study was to investigate anti-elastin antibodies of the IgG and IgM types in sera of patients suffering from lung cancer, using the DOT immunobinding assay. We studied 96 pathological and 40 control sera. Anti-elastin antibodies were found to be present in 45% of patients with small cell lung cancer, 19% of subjects with adenocarcinoma and not-identified lung tumor and 15% of patients with squamous cell lung cancer. They circulated in 5% of control persons only. The highest values of their titers were observed in the advanced stages of disease. In 55% of anti-elastin antibody positive small cell lung cancer patients, antibodies were of the IgM type, suggesting the initial step of the autoimmunization to elastin.

Topics: Adenocarcinoma; Antibodies, Neoplasm; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Elastin; Female; Humans; Immunoblotting; Immunoglobulin G; Immunoglobulin M; Lung Neoplasms; Male; Middle Aged

Topics: Collagen; Connective Tissue; Elastin; Evaluation Studies as Topic; Female; Humans; Hydroxylysine; Hydroxyproline; Lung Neoplasms; Male; Middle Aged; Neoplasm Proteins; Peptides

Interactions between the extracellular matrix macromolecules and tumor cells are critical in the process of metastasis formation. We show here that elastins (both mature insoluble elastin and a 75-kDa soluble peptide: K-elastin) adhere rapidly to two cell lines with high metastatic capacities: a metastatic lung carcinoma cell line (3LL-HM) and a human amelanotic melanoma cell line (A-2058); by contrast the low-metastatic Lewis lung carcinoma cell line variant as well as a rhabdomyosarcoma cell line with a low metastatic potential bind to elastins to a much lower extent. 3H-labelled K-elastin was used in order to study elastin--3LL-HM interaction. It was found to be saturable (2 ng 3H-labelled K-elastin/10(6) cells), with one class of high-affinity binding sites having Kd equal to 1.3 nM and 16,000 sites/cell. The binding of K-elastin to 3LL-HM cells at its receptor triggered several cell responses; (a) increase of intracellular Ca2+ concentration; (b) induction of 3LL-HM chemotaxis toward the K-elastin gradient; (c) stimulation of the adherence of mature insoluble elastin. In contrast to non-transformed cells such as fibroblasts and smooth muscle cells, the adhesion kinetics of insoluble elastin to 3LL-HM did not exhibit a lag period; the rapid binding of insoluble elastin to the tumor cells was followed by its slow detachment from the cells, which lasted for 6 h. 3LL-HM cells but not human skin fibroblasts were shown to secrete elastinolytic activity inhibitable by metal-chelating agents. In vivo studies were performed in order to evaluate the influence of K-elastin binding to 3LL-HM cells on their ability to form lung colonies in mice. It was shown that pretreatment of 10(4) 3LL-HM cells with 10 microM K-elastin and the simultaneous i.v. injection into mice of 750 micrograms K-elastin together with the highly metastatic cells was able to reduce the number of lung colonies by more than 70% after 12 days.

Topics: Adult; Amino Acid Sequence; Animals; Cell Adhesion; Chemotaxis; Elastin; Female; Fibroblasts; Humans; Kinetics; Lung Neoplasms; Mice; Mice, Nude; Molecular Sequence Data; Neoplasm Metastasis; Pancreatic Elastase; Receptors, Cell Surface; Tumor Cells, Cultured

To study the pulmonary structural remodeling in pulmonary lymphangiomyomatosis, electron microscopy and light and electron microscopic immunohistochemical observations for elastin and alpha 1-antitrypsin were performed on five open lung biopsy samples. Lung specimens showed emphysema-like changes in areas of abnormally accumulated smooth muscle cells. In the alveolar walls having accumulated smooth muscle cells, elastic fibers were decreased in number, disrupted, granular, and occasionally accumulated. Ultrastructurally, elastic fibers in areas of smooth muscle cell accumulation showed poorly outlined amorphous components and a few microfibrils, and occasionally showed electron-dense granular deposits in and around the amorphous components. Spiraling collagen fibrils were frequently found associated with these abnormal elastic fibers. Immunohistochemistry for elastin showed even staining of amorphous components of elastic fibers in the areas of smooth muscle cell accumulation. alpha 1-Antitrypsin was also detected evenly in amorphous components of elastic fibers in the areas of smooth muscle cell accumulation. It is proposed that the emphysema-like lesions of lymphangiomyomatosis are mediated by the degradation of elastic fibers, and these degraded elastic fibers are related to an imbalance of the elastase/alpha 1-antitrypsin system similar to the probable pathogenesis of emphysema.

Topics: Adult; alpha 1-Antitrypsin; Elastic Tissue; Elastin; Emphysema; Female; Humans; Lung Neoplasms; Lymphangiomyoma; Middle Aged

Elastin surrounds microvessels in the pulmonary circulation and may pose a barrier to the extravasation of metastatic tumor cells. We find that lung-colonizing murine melanoma cells produce an enzymatic activity that degrades elastin. In addition, the elastin fragments liberated by enzymatic digestion of insoluble elastin stimulate tumor cell chemotaxis. Chemotactic activity is associated with other forms of soluble elastin, including alpha-elastin and tropoelastin. Val-Gly-Val-Ala-Pro-Gly, a synthetic peptide that is a repeat sequence in the elastin molecule, also displayed tumor cell chemotactic activity. The ability to degrade elastin and to migrate in response to soluble elastin peptides is not a property of all tumor cells, but it is most commonly found associated with metastatic tumor cells that colonize pulmonary tissue. We postulate that the ability to migrate in response to elastin fragments may facilitate tumor cell invasion of elastin-rich pulmonary tissue.

Topics: Animals; Chemotactic Factors; Elastin; Lung Neoplasms; Pancreatic Elastase; Tropoelastin; Tumor Cells, Cultured

Extracellular matrix proteins and their proteolytic products have been shown to modulate cell motility. We have found that certain tumor cells display a chemotactic response to degradation products of the matrix protein elastin, and to an elastin-derived peptide, VGVAPG. The hexapeptide VGVAPG is a particularly potent chemotaxin for lung-colonizing Lewis lung carcinoma cells (line M27), with 5 nM VGVAPG eliciting maximal chemotactic response when assayed in 48-microwell chemotaxis chambers. Binding of the elastin-derived peptide to M27 cells was studied using a tyrosinated analog (Y-VGVAPG) to allow iodination. Scatchard analysis of [125I]Y-VGVAPG binding to viable M27 tumor cells at both 37 and 4 degrees C indicates the presence of a single class of high affinity binding sites. The dissociation constant obtained from these studies (2.7 X 10(-9) M) is equivalent to the concentration of VGVAPG required for chemotactic activity. The receptor molecule was identified as an Mr 59,000 species by covalent cross-linking of the radiolabeled ligand to the M27 tumor cell surface and subsequent analysis of the cross-linked material by electrophoresis and size-exclusion high performance liquid chromatography. These results suggest that M27 tumor cell chemotaxis to VGVAPG is initiated by high affinity binding of the peptide to a distinct cell surface receptor.

Topics: Animals; Chemotactic Factors; Cross-Linking Reagents; Elastin; Lung Neoplasms; Molecular Weight; Neoplasm Proteins; Oligopeptides; Receptors, Cell Surface; Tumor Cells, Cultured