heparitin-sulfate and Carcinoma--Squamous-Cell

The destruction of the epithelial basement membrane is widely regarded as a clear criterion for invasive malignant tumor growth. Since, however, defects in the basement membrane may also occur in non-invasive conditions, such as inflammatory and proliferative lesions, and since it has been shown that particularly in highly differentiated squamous cell carcinomas a continuous basement membrane is mimicked by the presence of isolated components, this criterion seems to be of minor value for the diagnosis of malignancy. Despite these drawbacks, the immunolocalization of basement membrane material may still be of differential diagnostic significance in certain situations. This holds particularly true for invasive (ductal) breast carcinomas, which usually completely lack a basement membrane. Accordingly, sclerosing adenosis can be distinguished from invasive carcinoma, as a distinction can be made between neoplastic (malignant) tubular formations and reactive lesions.

Topics: Basement Membrane; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; Collagen; Female; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Laminin; Neoplasm Invasiveness; Prognosis; Proteoglycans

Syndecans form a family of cell surface proteoglycans, which can interact with various effector molecules, such as extracellular matrix (ECM) molecules and growth factors. Syndecan-1 is the most extensively studied member of the syndecan family. It is found mainly in epithelial cells, but its expression is developmentally regulated during embryonic development. It has been shown to mediate cell adhesion to several ECM molecules, and to act as a coreceptor for fibroblast growth factors, potent angiogenic growth factors involved also in differentiation. Syndecan-1 expression is reduced during malignant transformation of various epithelia, and this loss correlates with the histological differentiation grade of squamous cell carcinomas, lacking from poorly differentiated tumours. In squamous cell carcinomas of the head and neck, positive syndecan-1 expression correlates with a more favourable prognosis. Recent experimental studies on the role of syndecan-1 in malignant transformation have shown that syndecan-1 expression is associated with the maintenance of epithelial morphology, anchorage-dependent growth and inhibition of invasiveness in vitro. This review will focus on the biological role of syndecan-1 in normal epithelial differentiation and malignant transformation.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Adhesion; Cell Transformation, Neoplastic; Growth Substances; Heparitin Sulfate; Humans; Membrane Glycoproteins; Proteoglycans; Syndecan-1; Syndecans

In this report, we used liquid chromatography-mass spectrometry and Western blotting to analyze the content and structure of glycosaminoglycans, glycolipids and selected proteins to compare differences between patient-matched normal and cancerous lung tissues obtained from lung cancer patients. The cancer tissue samples contained over twice as much chondroitin sulfate (CS)/dermatan sulfate (DS) as did the normal tissue samples, while the amount of heparan sulfate (HS) and hyaluronan (HA) in normal and cancer tissues were not significantly different. In HS, several minor disaccharide components, including NS6S, NS2S and 2S were significantly lower in cancer tissues, while the levels of major disaccharides, TriS, NS and 0S disaccharides were not significantly different in normal and cancer tissues. In regards to CS/DS, the level of 4S disaccharide (the major component of CS-type A and DS) decreased and the level of 6S disaccharide (the major component of CS- type C) increased in cancer tissues. We also compared the content and structure of GAGs in lung tissues from smoking and non-smoking patients. Analysis of the glycolipids showed all lipids present in these lung tissues, with the exception of sphingomyelin were higher in cancer tissues than in normal tissues. Western analysis showed that syndecan 1 and 2 proteoglycans displayed much higher expression in cancer tissue/biopsy samples. This investigation begins to provide an understanding of patho-physiological roles on glycosaminoglycans and glycolipids and might be useful in identifying potential biomarkers in lung cancer.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Chondroitin Sulfates; Chromatography, Liquid; Dermatan Sulfate; Disaccharides; Female; Glypicans; Heparitin Sulfate; Humans; Hyaluronic Acid; Lung Neoplasms; Male; Middle Aged; Retrospective Studies; Smoking; Syndecan-1; Tandem Mass Spectrometry

Syndecan-1 binds to various extracellular matrix components via its heparan sulfate glycosaminoglycans (HS-GAG) and most of its biological functions are considered to be associated with this process. The aims of this study are to investigate its expression in cervical neoplasms.. We investigated the expression of both the syndecan-1 core protein and cell-surface HS-GAG by immunohistochemistry in 53 cervical intraepithelial neoplasm (CIN), 19 microinvasive, 143 invasive cervical cancers, and 29 metastatic lymph node samples, and analyzed correlations with various clinicopathological features.. The progression of CIN to early invasive cancer was found to associate with reduced levels of both syndecan-1 and HS-GAG expression. In squamous cell carcinomas, HS-GAG expression was significantly lower in cases with lymph-vascular space invasion. Additionally, the overall survival rates for patients exhibiting low HS-GAG expression was significantly lower than patients exhibiting high HS-GAG expression (P = 0.019). Low HS-GAG expression in positive nodes was determined to be a disease-free and overall survival prognostic factor (P = 0.028 and P = 0.018, respectively).. The loss of syndecan-1 and HS-GAG expression is an early event in cervical carcinogenesis. The loss of HS-GAG expression particularly in positive nodes can serve as an indicator of aggressive disease potential and poor prognosis in patients with invasive cervical cancer.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Female; Heparitin Sulfate; Humans; Immunohistochemistry; Lymph Nodes; Lymphatic Metastasis; Membrane Glycoproteins; Middle Aged; Neoplasm Invasiveness; Prognosis; Proteoglycans; Syndecan-1; Syndecans; Treatment Outcome; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

In previous studies, we have demonstrated that human heparanase (endo-beta-D-glucuronidase) is localized primarily in a perinuclear pattern within lysosomes and late endosomes, and occasionally may be surface associated and secreted. The presence of two potential nuclear localization sequences in human heparanase, led us to investigate heparanase translocation into the nucleus and subsequent degradation of nuclear heparan sulfate. Applying cell fractionation, Western blot analysis, determination of heparanase activity and confocal microscopy, we identified heparanase within the nuclei of human glioma and breast carcinoma cells and estimated its amount to be about 7% of the cytosolic enzyme. Our results indicate that nuclear heparanase colocalizes with nuclear heparan sulfate and is enzymaticaly active. Moreover, following uptake of latent 65 kDa heparanase by cells that do not express the enzyme, an active 50 kDa heparanase was detected in the cell nucleus, capable of degrading both nuclear and extracellular matrix-derived heparan sulfate. Immunohistochemical examination of human squamous cell carcinoma specimens revealed a prominent granular staining of heparanase within the nuclei of the epithelial tumor cells vs no nuclear staining in the adjacent stromal cells. Taken together, it appears that heparanase is translocated into the cell nucleus where it may degrade the nuclear heparan sulfate and thereby affect nuclear functions that are thought to be regulated by heparan sulfate. Nuclear localization of heparanase suggests that the enzyme may fulfill nontraditional functions (ie, regulation of gene expression and signal transduction) apart of its well-documented involvement in cancer metastasis, angiogenesis and inflammation.

Topics: Active Transport, Cell Nucleus; Amino Acid Sequence; Animals; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Nucleus; Glucuronidase; Heparin; Heparitin Sulfate; Humans; Molecular Sequence Data; Mouth Neoplasms; Nuclear Localization Signals; Rats; Recombinant Proteins; Substrate Specificity; Transfection

It is still unknown how jaw bone remodeling occurs at actual invasion sites of oral squamous cell carcinomas. Since there is no other human carcinomas which make a direct invasion of the bone, gingival carcinomas are valuable examples.. Twelve surgical specimens of gingival squamous cell carcinoma were examined histopathologically and immunohistochemically for remodeling of bone and its surrounding tissue.. Three types of bone interfaces with carcinomatous invasion were distinguished. These included areas with bone resorption, smooth bone surface and new bone formation. In the bone-resorption area, numerous osteoclasts were located along the bone surface, which was surrounded by myxoid stroma. The myxoid stroma was characterized by immunopositivity for heparan sulfate proteoglycan (HSPG), abundant vascularity and macrophagic infiltration. In the bone-formation area, rows of osteoblasts were aligned on the bone surface. The stroma around osteoblasts was also HSPG-immunopositive, poor in vascularity but rich in activated fibroblasts. In the smooth-bone area, the stroma showed an organizing phase of granulation tissue with slender fibroblasts and mature collagen fibers but with less vascularity and inflammatory infiltrates.. The results indicate that the stromal architecture, especially in terms of its inflammatory cellular, vascular and matrix compositions, is strictly regulated in the timing and site of jaw bone remodeling which is causes by carcinomatous invasion.

Topics: Antigens, CD; Antigens, Differentiation, Myelomonocytic; Bone Remodeling; Bone Resorption; Carcinoma, Squamous Cell; Cathepsin K; Cathepsins; Gingival Neoplasms; Heparitin Sulfate; Humans; Immunoenzyme Techniques; Jaw; Neoplasm Invasiveness; Neovascularization, Pathologic; Osteoclasts; Tenascin

Phage display was used to identify homing peptides for blood vessels in a mouse model of HPV16-induced epidermal carcinogenesis. One peptide, CSRPRRSEC, recognized the neovasculature in dysplastic skin but not in carcinomas. Two other peptides, with the sequences CGKRK and CDTRL, preferentially homed to neovasculature in tumors and, to a lesser extent, premalignant dysplasias. The peptides did not home to vessels in normal skin, other normal organs, or the stages in pancreatic islet carcinogenesis in another mouse model. The CGKRK peptide may recognize heparan sulfates in tumor vessels. The dysplasia-homing peptide is identical to a loop in kallikrein-9 and may bind a kallikrein inhibitor or substrate. Thus, characteristics of the angiogenic vasculature distinguish premalignant and malignant stages of skin tumorigenesis.

Topics: Animals; Blood Vessels; Carcinoma, Squamous Cell; Ectodermal Dysplasia; Heparitin Sulfate; Immunohistochemistry; Kallikreins; Mice; Neoplasm Proteins; Neoplasm Staging; Neovascularization, Pathologic; Peptide Library; Peptides; Skin

Previous in vitro studies have shown CD44 isoforms containing the alternatively spliced exon v3 (CD44v3) to be modified with heparan sulphate (HS) and to bind HS-binding basic fibroblast growth factor (bFGF). Here, we demonstrate that exogenously added bFGF is also bound in vivo by CD44v3-positive keratinocytes in normal skin and by tumour cells in basal cell carcinoma and squamous cell carcinoma (SCC), two skin cancers of keratinocyte origin. bFGF binding and CD44v3 expression were colocalized in cultured human normal keratinocytes (HNK) and on the SCC cell line A431. By contrast, benign or malignant tumours of melanocyte origin failed to express CD44v3 and bound no bFGF. The bFGF binding to normal or transformed keratinocytes in vivo and in vitro was dependent on HS modification, as it was completely eliminated by pretreatment with heparitinase or by blocking with free heparin, whereas chondroitinase had no effect. In addition, specific removal of CD44v3 by antibody-induced shedding also diminished bFGF binding to keratinocytes. Furthermore, bFGF stimulated the proliferation of CD44v3-positive HNK and A431 in a dose-dependent fashion. This bFGF effect was again completely abolished by heparitinase or free heparin, but not by chondroitinase. In aggregate, our results suggest that a function of HS-modified CD44 isoforms such as CD44v3 in skin is to present the HS-binding growth factor bFGF, thereby stimulating the proliferation of normal or transformed keratinocytes.

Topics: Antigens, Neoplasm; Carcinoma, Squamous Cell; Cell Culture Techniques; Cell Division; Fibroblast Growth Factor 2; Heparitin Sulfate; Humans; Hyaluronan Receptors; Infant, Newborn; Keratinocytes; Male; Skin; Skin Neoplasms; Tumor Cells, Cultured

Heparan sulfate (HS) functions as a co-factor in several signal-transduction systems that affect cellular growth, differentiation, adhesion and motility. HS, therefore, may also play a role in the malignant transformation of cells, tumor growth, cell invasiveness and the formation of tumor metastases. To explore this hypothesis, we analyzed the expression of HS and heparan sulfate proteoglycan (HSPG) in histological sections of human lung-cancer tissues and assayed for the presence of HSPGs in extracts of human lung-cancer cell lines, using a panel of native HS-, delta-HS- and HSPG (syndecan, glypican, CD44 and perlecan) core protein-specific monoclonal antibodies. Compared to normal epithelia, non-small-cell lung carcinomas, particularly poorly differentiated tumors, often expressed reduced amounts of the major cell surface-associated HSPGs (most consistently of syndecan-1). CD44 or CD44-variant proteins, in contrast, were found on all tumor cells, irrespective of their differentiation. Perlecan, a matrix-associated HSPG found in the basement membrane of normal bronchial epithelium, was consistently undetectable in invasive bronchogenic carcinomas. Staining reactions for native HS were consistently reduced in squamous-cell lung carcinomas, in the cells in contact with the stroma and in the less differentiated areas of these tumors. Reactions for delta-HS, however, were not reduced, suggesting a structural change in the HS of these tumor cells. Poorly differentiated adenocarcinomas, in contrast, yielded strong HS and delta-HS reactions. Marked differences in HSPG expression also were observed among various non-small-cell lung carcinoma cell lines. Our results suggest that poorly differentiated lung tumors have markedly altered patterns of HSPG expression, which may contribute to their invasive phenotype.

Topics: Adenocarcinoma; Carcinoma, Squamous Cell; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Hyaluronan Receptors; Lung Neoplasms; Lymphatic Metastasis; Mediastinum; Neoplasm Proteins; Proteoglycans; Tumor Cells, Cultured

Epithelial structures are separated from the stroma by a basement membrane (BM) which serves as a barrier for the epithelial cells. Invasive tumour growth as in laryngeal carcinoma, involves the degradation of this barrier. In our present immunohistochemical analysis, we evaluated both the quantitative and the qualitative changes in the BM composition of 50 laryngeal carcinomas. This analysis comprised the localisation of the BM components collagen IV, laminin, and fibronectin. Furthermore, we applied antibodies against BM components that have not, or only very rarely, been analysed in malignant squamous cell neoplasms--particularly collagen VII and heparan sulfate proteoglycan (HSPG). In this study we provide considerable evidence that varying amounts of preserved BM material can be found in laryngeal carcinoma of different grades of tumour differentiation. Especially by comparison of the different staining patterns for collagen IV and collagen VII, we recognised far more gaps in the staining of the BM by collagen VII than by collagen IV. This fact is underlined by the observation that even in G1 carcinomas collagen VII showed in almost 40% of the cases a complete loss of BM staining. In general, we found a correlation between the amount of preserved BM deposition and the grade of tumour differentiation. This fact may underline the significance of the immunohistochemically detectable amount of BM components as a prognostically relevant parameter.

Topics: Adult; Aged; Basement Membrane; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Differentiation; Cell Division; Cell Transformation, Neoplastic; Collagen; Female; Fibronectins; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Immunoenzyme Techniques; Laminin; Laryngeal Neoplasms; Male; Middle Aged; Neoplasm Invasiveness; Proteoglycans

The basement lamina of the epithelium of the true vocal cords of 34 inpatients suffering from chronic laryngitis, Reinke's oedema and squamous cell carcinoma have been investigated with the electron and immunofluorescence microscope. In chronic laryngitis, the lamina fibroreticularis is thickened (due to collagen type VII), corresponding to the clinical finding. In this layer mobile cells of the connective tissue can be found. In cases of Reinke's oedema it is the lamina densa which might be thickened. In this disease, also lamina densa-like material can protrude into the lamina fibroreticularis, and the number of anchoring filaments is increased. In squamous cell carcinoma we found the basement lamina irregularly arranged and folded. The lamina densa was always interrupted by numerous small gaps and in some areas the basement membrane could not be identified over a long distance. Lamina densa-like material was also found between the tumour cells within the epithelium. With the immunofluorescence microscope this material was proven as laminin, collagen type IV and heparan sulfate proteoglycan. Our investigation shows that malignant as well as benign lesions of the true vocal cords are characterised by distinct fine structural findings even concerning the basement lamina.

Topics: Adult; Aged; Basement Membrane; Biomarkers, Tumor; Carcinoma, Squamous Cell; Collagen; Epithelium; Female; Fibronectins; Fluorescent Antibody Technique; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Hoarseness; Humans; Laryngeal Edema; Laryngeal Neoplasms; Laryngitis; Larynx; Male; Microscopy, Electron; Middle Aged; Neoplasm Invasiveness; Proteoglycans; Vocal Cords

Topics: Antibodies, Monoclonal; Basement Membrane; Carcinoma, Squamous Cell; Chondroitin Sulfates; Chondroitinases and Chondroitin Lyases; Collagen; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Immunohistochemistry; Keratosis; Laminin; Proteoglycans; Skin Neoplasms

Topics: Carcinoma, Squamous Cell; Dermatan Sulfate; Diabetes Mellitus, Type 1; Female; Glycosaminoglycans; Heparitin Sulfate; Humans; Hyaluronic Acid; Keratan Sulfate; Lung Neoplasms; Male; Middle Aged; Protein Binding

An immunohistochemical study of samples from 12 cervical carcinomas was carried out using monoclonal antibodies against laminin (L) and heparan sulfate proteoglycan (HSP). Cryostat sections of operative and biopsy specimens were processed by the immunoperoxidase method. L and HSP were identified in the basal membrane of normal and neoplastic epithelium. Basal membrane disruptions were found in in situ, microinvasive and invasive carcinomas. However, in the first group they were less common and occurred only in the presence of lymphohistiocytic infiltration of the underlying stroma. An inverse relationship was established between depth of invasion and amount of L and HSP in the basal membrane of tumor epithelium.

Topics: Adult; Carcinoma, Squamous Cell; Female; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Immunoenzyme Techniques; Laminin; Middle Aged; Neoplasm Invasiveness; Proteoglycans; Uterine Cervical Neoplasms

To investigate alterations in the basement membrane (BM) in squamous cell carcinoma (SCC), we investigated 20 tumors. Four had the cytologic characteristics of Bowen's disease (SCC-BD) and 16 did not have them (SCC-NB). Tumors were studied immunohistochemically by double immunofluorescent staining by using mouse monoclonal antibodies to the core protein of heparan sulfate proteoglycan (HSPG) and chondroitin 6-sulfate glycosaminoglycan (Ch6S) as well as rabbit antiserum to laminin (LN) and type IV collagen (C-4). In well-differentiated and highly keratinized SCC-NB, LN, C-4, and HSPG could be detected in the tumor nest BM and showed no loss of continuity, but they were largely lost in poorly differentiated and poorly keratinized SCC-NB. This suggests that poorly differentiated SCC-NB cause greater enzymatic degradation of BM components than well-differentiated SCC-NB. Ch6S was detected in parts of the BM of SCC-BD, but it was absent in all SCC-NB examined. It appears that SCC-NB have lost the ability to synthesize Ch6S, and that SCC-BD degrade Ch6S although they continue to produce it. Thus, it appears that in SCC the BM is qualitatively different from that of normal epidermis, and that SCC-BD can be distinguished from SCC-NB by the Ch6S content of the BM.

Topics: Antibodies; Antibodies, Monoclonal; Basement Membrane; Bowen's Disease; Carcinoma, Squamous Cell; Chondroitin Sulfate Proteoglycans; Collagen; Fluorescent Antibody Technique; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Laminin; Reference Values; Skin; Skin Neoplasms

The glycosaminoglycans of four normal human bladders and fourteen bladder cancers were characterized and quantitated (after proteolytic extraction) by specific enzyme digestion, cellulose acetate electrophoresis and densitometry. Hyaluronic acid, heparan sulfate, dermatan sulfate and chondroitin sulfate were identified in both normal and cancerous bladders. Hyaluronic acid and dermatan sulfate were the major glycosaminoglycans of the normal epithelium/submucosa while heparan sulfate and dermatan sulfate were predominant in normal bladder muscle. Bladder cancer glycosaminoglycan content was influenced by the stage and grade of the neoplasm. Hyaluronic acid and dermatan sulfate tended to decrease and chondroitin sulfate to increase in infiltrating cancers, whereas a decrease in the percentage of heparan sulfate correlated closely with higher grade tumors. The bladder cancer glycosaminoglycan profile may be indicative of the tumor's invasive potential.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Child; Chondroitin Sulfates; Densitometry; Electrophoresis, Cellulose Acetate; Female; Glycosaminoglycans; Heparitin Sulfate; Humans; Hyaluronic Acid; Male; Middle Aged; Urinary Bladder; Urinary Bladder Neoplasms

The quantitative changes of glycosaminoglycans in tumor tissue of human lung cancers (2 squamous cell carcinomas, 4 adenocarcinomas and 5 small cell carcinomas) were studied. The total amount of glycosaminoglycans in human lung cancer tissues increased 1.4 to 4 times in comparison with that in normal lung tissues. The increase in tissue content of glycosaminoglycans was accompanied by an increase in the chondroitin sulfate level in every histologic type of lung cancer, as well as by a marked increase in hyaluronic acid level in squamous cell carcinomas, and a moderate increase in its level in small cell carcinomas. The concentrations of dermatan sulfate and heparan sulfate in lung cancer tissues did not show any significant changes compared with those in normal lung tissues. The increase in total amount and changes in the composition of glycosaminoglycans in human lung cancer tissue were closely related to the histologic type of the tumor.

Topics: Adenocarcinoma; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Chondroitin Sulfates; Dermatan Sulfate; Electrophoresis, Cellulose Acetate; Glycosaminoglycans; Heparitin Sulfate; Humans; Hyaluronic Acid; Lung Neoplasms

The glycosaminoglycans were prepared by exhaustive Pronase digestion and alkaline treatment of squamous cell carcinoma and adenocarcinoma tissues of human lung, and of tissues taken at a site distant from the tumor as a control. The glycosaminoglycan classes were characterized by chemical enzymic, and electrophoretic methods. The presence of oversulfated chondroitin-and/or dermatan-sulfates which have not up till now been found in lung tissues was also demonstrated in carcinoma and control tissues, their contents being higher in the carcinoma tissues. The levels of whole glycosaminoglycans were markedly increased in carcinoma tissue. The classes of glycosaminoglycans which increased in lung carcinoma tissue were predominatly chondroitin-4-and/or-6-sulfates and hyauronic acid.

Topics: Adenocarcinoma; Carcinoma, Squamous Cell; Chondroitin Sulfates; Glycopeptides; Glycosaminoglycans; Heparitin Sulfate; Hexosamines; Humans; Hyaluronic Acid; Lung; Lung Neoplasms; Sulfates