heparitin-sulfate and Cell-Transformation--Neoplastic

Proteoglycans are macromolecules formed by a protein core to which sugar chains are covalently attached. They are present on the cell surface and in the ECM of living things. In normal liver syndecan-1 is the dominant transmembrane proteoglycan, trace amounts of ECM proteoglycans are in the stromal components. The amounts of proteoglycans we studied increase in liver cirrhosis. In liver cancer abnormal localization of syndecan-1 and stroma rich in agrin was characteristic. The core proteins as well as the sugar chains of proteoglycans interact with and modulate the effect of regulatory factors. This implies that structural alterations of proteoglycans contribute to the development of malignant phenotype. Heparan sulfate chains of liver cancer are undersulfated with decreased or altered biological activity. Their binding capacity for transcription factor decreases, and they do not inhibit topoisomerase I enzyme. Truncated form of syndecan-1 lacking the extracellular domain of the molecule induces differentiation of hepatoma cell line and inhibits the shedding of syndecan-1. This phenomenon calls attention to the importance of syndecan-1 shedding in the regulation of cell behavior.

Topics: Animals; Cell Differentiation; Cell Transformation, Neoplastic; Heparitin Sulfate; Humans; Liver; Liver Cirrhosis; Liver Diseases; Liver Neoplasms; Membrane Glycoproteins; Proteoglycans; Signal Transduction; Syndecan-1; Syndecans

Topics: Animals; Apoptosis; Carrier Proteins; Cell Transformation, Neoplastic; Endoplasmic Reticulum; Gene Expression Regulation, Developmental; Golgi Apparatus; Heparitin Sulfate; Humans; Intercellular Signaling Peptides and Proteins; Neoplasms; Nervous System; Proteins; Signal Transduction; Sulfatases; Wnt Proteins

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

Cell surface proteoglycans are strategically positioned to regulate interactions between cells and their surrounding environment. Such interactions play key roles in several biological processes, such as cell recognition, adhesion, migration, and growth. These biological functions are in turn necessary for the maintenance of differentiated phenotype and for normal and neoplastic development. There is ample evidence that a special type of proteoglycan bearing heparan sulfate side chains is localized at the cell surface in a variety of epithelial and mesenchymal cells. This molecule exhibits selective patterns of reactivity with various constituents of the extracellular matrix and plasma membrane, and can act as growth modulator or as a receptor. Certainly, during cell division, membrane constituents undergo profound rearrangement, and proteoglycans may be intimately involved in such processes. The present work will focus on recent advances in our understanding of these complex macromolecules and will attempt to elucidate the biosynthesis, the structural diversity, the modes of cell surface association, and the turnover of heparan sulfate proteoglycans in various cell systems. It will then review the multiple proposed roles of this molecule, with particular emphasis on the binding properties and the interactions with various intracellular and extracellular elements. Finally, it will focus on the alterations associated with the neoplastic phenotype and will discuss the possible consequences that heparan sulfate may have on the growth of normal and transformed cells.

Topics: Cell Communication; Cell Division; Cell Transformation, Neoplastic; Chondroitin Sulfate Proteoglycans; Cytoskeleton; Glycosaminoglycans; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Membrane Glycoproteins; Phenotype; Proteoglycans

There is a growing realization that the whole tumor cell-matrix complex must be investigated in order to fully understand the process of cancer growth and metastasis. Proteoglycans are intrinsic constituents of the cell surface, extracellular matrix, and basement membrane, three logistically and functionally important structures involved in most cellular interactions. Proteoglycans influence the behavior of normal and malignant cells by virtue of their expanded configuration, polyanionic nature and, most of all, by their ability to interact with a variety of cellular products. Consequently, they have been implicated in a number of biological processes including proliferation, recognition, adhesion, and migration. They can serve as links between the extracellular and intracellular environment and thus transduce key biological signals. They can act as receptors for interstitial collagens and other matrix proteins and thus contribute to the organization of pericellular matrix. During neoplastic development there is a profound structural rearrangement of these macromolecules at both the plasma membrane and the pericellular level. Qualitative and quantitative abnormalities in proteoglycan metabolism may contribute to the establishment of some well-known neoplastic properties, including lack of cohesiveness, abnormal assembly of extracellular matrix, abnormal growth, and invasion. The present work will focus on recent advances in our understanding of these complex macromolecules and on some of the alterations associated with the neoplastic phenotype, and will then attempt to elucidate some of the mechanisms regulating these changes.

Topics: Biological Products; Cell Division; Cell Transformation, Neoplastic; Cytokines; Glycosaminoglycans; Heparitin Sulfate; Humans; Neoplasms; Proteoglycans

The central question in cell biology is how cells detect, interact and respond to extracellular matrix. The cell surface molecules, which mediate this recognition, consist of a lipophilic membrane domain and an ectodomain binding matrix materials. One group of this kind of molecules is the cell surface heparan sulfate proteoglycans (HSPG). This review summarizes recent information obtained on the cell surface PG of mouse mammary epithelial cells. The glycosaminoglycan containing ectodomain of this PG binds with high affinity Type I, III and V collagen fibrils and the C-terminal heparin binding domain of fibronectin. The PG is mobile on the cell surface, but can be immobilised by ligand binding. At the same time the PG associates with cytoskeleton and links the epithelial cytoskeleton to extracellular matrix. Thus the PG can mediate the changes in the matrix into changes in cellular behaviour, often seen during the regulation of cell shape, proliferation and differentiation. The cell surface PG is also released from the cell surface by cleaving the matrix-binding ectodomain from the membrane domain. Because of the binding properties of the ectodomain, this shedding may provide a means by which epithelial cells loosen their association with the matrix and with other cells, e.g., during normal epithelial development and the invasion of carcinomas.

Topics: Animals; Cell Membrane; Cell Transformation, Neoplastic; Chondroitin Sulfate Proteoglycans; Cytoskeleton; Epithelium; Extracellular Matrix; Glycosaminoglycans; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Proteoglycans

Basement membranes are ubiquitous tissue constituents which occur as supportive structure adjacent to epithelium, endothelium, mesothelium and also around smooth as well as striated muscle cells, Schwann cells and fat cells. In various types of cancer, basement membranes have been extensively studied by electron microscopy. Often basement membrane interruptions were seen in invasive neoplasms but in some tumors the neoplastic cells were surrounded by a continuous basal lamina. Recent immunocytochemical studies have shown that in invasive carcinomas the neoplastic cells often lack a continuous basement membrane. This may be caused by catabolic activity of invasive tumor cells, which have been shown to produce specific collagenases, or by insufficient production and/or extracellular assembly of basement membrane components by the neoplastic epithelial cells. In diagnostic histopathology, immunocytochemical staining of basement membrane components such as type IV collagen and laminin may help to distinguish between noninvasive (benign or in situ) and invasive lesions. Furthermore, in carcinomas the extent of the expression of basement membrane components may be correlated with the degree of differentiation of the neoplastic cells. Finally, in soft tissue tumors, basement membrane staining may be helpful for the differentiation of basement membrane producing neoplasms (e.g. of vascular, neural, smooth muscle or striated muscle origin) from non-basement membrane producing neoplasms (e.g. of fibroblastic origin).

Topics: Basement Membrane; Cell Differentiation; Cell Transformation, Neoplastic; Chondroitin Sulfate Proteoglycans; Collagen; Fluorescent Antibody Technique; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Immunoenzyme Techniques; Laminin; Microscopy, Electron; Neoplasm Invasiveness; Neoplasms

Proteoglycans have been studied to a limited extent in lymphoid cells. In this study we have investigated the expression of proteoglycans in B-cells, CD4+ T-cells, CD8+ T-cells, natural killer cells, as well as in nine different cell lines established from patients with lymphoid malignancies. Serglycin was the major proteoglycan expressed at mRNA level by the primary lymphocytes. None of the syndecans or glycpicans was detected at mRNA level in the primary lymphocytes, except for syndecan-4 in CD4+ T-cells and CD8+ T-cells. All lymphoid cell lines expressed serglycin mRNA, as well as one or several members of the syndecan and glypican families. Further, increased synthesis of proteoglycans was found in the cell lines compared to the primary lymphocytes, as well as the presence of heparan sulfate on the cell surface of five of the cells lines. Western blot analysis showed a close correlation between serglycin mRNA level and expression of serglycin core protein. Our results show that serglycin is a major proteoglycan in all the normal lymphoid cells and that these cells carry little, or none, proteoglycans on the cell surface. Serglycin was also a major proteoglycan in the malignant lymphoid cells, but these also expressed one or more types of cell surface proteoglycans. Thus, malignant transformation of lymphoid cells may be followed by increased synthesis of proteoglycans and expression of cell surface proteoglycans.

Topics: Cell Line, Tumor; Cell Transformation, Neoplastic; Hematologic Neoplasms; Heparitin Sulfate; Humans; Lymphocytes; Neoplasm Proteins; Proteoglycans; RNA, Messenger; RNA, Neoplasm; Syndecan-4; Vesicular Transport Proteins

Basic fibroblast growth factor (FGF-2) and its respective tyrosine kinase receptors, form an autocrine loop that affects human melanoma growth and metastasis. The aim of the present study was to examine the possible participation of various glycosaminoglycans, i.e. chondroitin sulfate, dermatan sulfate and heparin on basal and FGF-2-induced growth of WM9 and M5 human metastatic melanoma cells. Exogenous glycosaminoglycans mildly inhibited WM9 cell's proliferation, which was abolished by FGF-2. Treatment with the specific inhibitor of the glycosaminoglycan sulfation, sodium chlorate, demonstrated that endogenous glycosaminoglycan/proteoglycan production is required for both basal and stimulated by FGF-2 proliferation of these cells. Heparin capably restored their growth, and unexpectedly exogenous chondroitin sulfate to WM9 and both chondroitin sulfate and dermatan sulfate to M5 cells allowed FGF-2 mitogenic stimulation. Furthermore, in WM9 cells the degradation of membrane-bound chondroitin/dermatan sulfate stimulates basal growth and even enhances FGF-2 stimulation. The specific tyrosine kinase inhibitor, genistein completely blocked the effects of FGF-2 and glycosaminoglycans on melanoma proliferation whereas the use of the neutralizing antibody for FGF-2 showed that the mitogenic effect of chondroitin sulfate involves the interaction of FGF-2 with its receptors. Both the amounts of chondroitin/dermatan/heparan sulfate and their sulfation levels differed between the cell lines and were distinctly modulated by FGF-2. In this study, we show that chondroitin/dermatan sulfate-containing proteoglycans, likely in cooperation with heparan sulfate, participate in metastatic melanoma cell FGF-2-induced mitogenic response, which represents a novel finding and establishes the central role of sulfated glycosaminoglycans on melanoma growth.

Topics: Autocrine Communication; Cell Differentiation; Cell Line, Tumor; Cell Proliferation; Cell Transformation, Neoplastic; Chondroitin Sulfates; Fibroblast Growth Factors; Glycosaminoglycans; Heparitin Sulfate; Humans; Melanoma; Neoplasm Metastasis; Protein-Tyrosine Kinases; Proteoglycans

Heparan sulfate inhibits the proliferation of normal human lung fibroblasts (HFL-1) but not of a human lung carcinoma cell-line (A549). In this study we investigated possible mechanisms and structural requirements by which antiproliferative heparan sulfates exerts its effects on binding, uptake and subcellular localisation. Both HFL-1 and A549 cells were incubated with 125I- or rhodamine-labeled L-iduronate-rich antiproliferative heparan sulfate species as well as L-iduronate-poor inactive ones. The antiproliferative heparan sulfate was bound to the cell surface on both HFL-1 and A549 cells, but to a lesser extent and with less affinity to A549 cells. Both cell types bound the antiproliferative heparan sulfate with one high- and with one low affinity site. The L-iduronate-poor heparan sulfate bound to a lesser extent and with less affinity to both cell types compared to the antiproliferative heparan sulfate. The antiproliferative heparan sulfate accumulated in the cytoplasm of HFL-1 cells after 24 h incubation, but after 72 h it was found evenly distributed in the nucleus. The time-scale for antiproliferative activity correlated with nuclear localization. In contrast, in A549 cells it was only found near the nuclear membrane. The inactive heparan sulfate was taken up in considerably smaller amounts compared to the antiproliferative heparan sulfate and could not be detected in the nucleus of either HFL-1 or A549 cells. Our data suggest that the antiproliferative activity of L-iduronate-rich heparan sulfate on normal fibroblasts may be due to direct effects on nuclear processes, such as gene transcription.

Topics: Animals; Binding Sites; Cattle; Cell Nucleus; Cell Transformation, Neoplastic; Cells, Cultured; Fibroblasts; Growth Inhibitors; Heparitin Sulfate; Humans; Lung; Lung Neoplasms; Microscopy, Confocal; Radioligand Assay; Tumor Cells, Cultured

Heparan sulfate at cell surfaces and in the extracellular matrix regulates cell proliferation and adhesion by binding to growth factors and matrix proteins via structurally specific oligosaccharide domains. We have used the hormonally regulated mouse mammary carcinoma cell line S115 as a model to elucidate the effect of malignant transformation on the structure of heparan sulfate. When cultured in the presence of testosterone, S115 cells form tumor cell colonies in soft agar and exhibit fibroblast-like morphology; withdrawal of testosterone results in a loss of the tumorigenic capacity and a switch towards epithelial morphology. Metabolically 35SO4-labeled heparan sulfate was isolated from testosterone-treated and non-treated S115 cells and subjected to structural analysis. We found that the testosterone-dependent malignant transformation was associated with reduced sulfation of heparan sulfate due to a approximately 40% decrease in the amount of GlcN6S units. By contrast, no significant differences were observed in the amounts of 2-O-sulfate or N-sulfate groups. The reduced 6-O-sulfation of GlcN units in heparan sulfate from transformed S115 cells led to a marked decrease in the amount of trisulfated IdoA2S-GlcNS6S units (IdoA, L-iduronic acid), implicated in many heparan sulfate-protein interactions.

Topics: Animals; Cell Transformation, Neoplastic; Chromatography, Gel; Chromatography, Ion Exchange; Disaccharides; Epithelial Cells; Female; Heparitin Sulfate; Mammary Glands, Animal; Membrane Glycoproteins; Mice; Models, Biological; Proteoglycans; Recombinant Proteins; Sulfates; Sulfur Radioisotopes; Syndecans; Testosterone; Transfection; Tumor Cells, Cultured

Syndecans are a four member family of cell surface proteoglycans, which via their heparan sulfate chains can bind both extracellular matrix molecules and heparin-binding growth factors. In signal transduction they cooperate with tyrosine kinase receptors and thus can participate in the regulation of cell growth and behaviour. Besides the fact that their developmental expression follows morphogenetic rather than histological boundaries, also changes in their expression take place during development of diseases. Among these, the loss of syndecan expression during malignant transformation suggests that syndecans could play a role in restricting tumor growth and metastasis.

Topics: Awards and Prizes; Cell Division; Cell Transformation, Neoplastic; Extracellular Matrix Proteins; Fibroblast Growth Factor 1; Heparitin Sulfate; Membrane Glycoproteins; Molecular Biology; Proteoglycans; Signal Transduction; Sweden; Syndecans

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

F9 embryonal cells can be induced to differentiate and synthesize basement membrane proteins. Perlecan and laminin are two basement membrane constituents that have extensive regions of homology. Expression of perlecan and laminin B1 genes was followed during differentiation of F9 cells by measurements of transcription rate and mRNA abundance using nuclear run on assays and Northern hybridizations, respectively. The rate of precursor protein synthesis was determined by immunoprecipitation from lysates of pulse-labeled F9 cells. The results showed that perlecan gene expression responds more rapidly after induction than does laminin B1 gene expression but is ultimately expressed at a substantially lower level than laminin. Thus, the perlecan and laminin genes appear to be regulated by different mechanisms and their gene products are not made in stoichiometric amounts.

Topics: Animals; Blotting, Northern; Bucladesine; Carcinoma, Embryonal; Cell Transformation, Neoplastic; Gene Expression; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Laminin; Mice; Proteoglycans; RNA, Messenger; Transcription, Genetic; Tretinoin; Tumor Cells, Cultured

Topics: Cell Transformation, Neoplastic; Cells, Cultured; Child; Chromatography, High Pressure Liquid; Epidermis; Fibronectins; Glycosaminoglycans; Heparitin Sulfate; Humans; Infant, Newborn; Keratins; Male; Sulfates

The effects of glycosaminoglycans (GAGs), especially heparan sulfate, on human and porcine vascular endothelial cell growth in the presence of 10% fetal bovine serum (FBS) or so-called heparin-binding growth factors, acidic FGF (aFGF) or basic FGF (bFGF), were investigated in a series of studies in order to clarify their role in cell proliferation. The promotion of normal cell growth by aFGF was suppressed by heparan sulfate but enhanced by heparin, while growth promotion by bFGF was suppressed by both GAGs. Chondroitin sulfate and hyaluronic acid showed no significant suppression of promotion, even at much higher concentrations. The growth of spontaneously transformed cells was enhanced by heparan sulfate or heparin in the presence of 10% FBS or aFGF, while growth promotion in the presence of bFGF was suppressed by both GAGs. From these results, we concluded that heparan sulfate and heparin are not always functional analogs and that in some cases they exert opposite actions on the modulation of normal endothelial cell growth. The findings that heparan sulfate enhanced the growth of transformed cells but suppressed the growth of normal cells in the presence of 10% FBS or aFGF were consistent with those of our previous studies on human fibroblasts, confirming the occurrence of some common alterations in the signal transduction system or cell surface upon cellular transformation. The role of heparan sulfate in the modulation of endothelial cell proliferation in vivo is also discussed.

Topics: Cell Division; Cell Line; Cell Transformation, Neoplastic; Endothelium, Vascular; Female; Fibroblast Growth Factors; Glycosaminoglycans; Heparin; Heparitin Sulfate; Humans; Umbilical Veins

Basement-membrane proteoglycans, biosynthetically labelled with [35S]sulphate, were isolated from normal and transformed mouse mammary epithelial cells. Proteoglycans synthesized by normal cells contained mainly heparan sulphate and, in addition, small amounts of chondroitin sulphate chains, whereas transformed cells synthesized a relatively higher proportion of chondroitin sulphate. Polysaccharide chains from transformed cells were of lower average Mr and of lower anionic charge density compared with chains isolated from the untransformed counterparts, confirming results reported previously [David & Van den Berghe (1983) J. Biol. Chem. 258, 7338-7344]. A large proportion of the chains isolated from normal cells bound with high affinity to immobilized antithrombin, and the presence of 3-O-sulphated glucosamine residues, previously identified as unique markers for the antithrombin-binding region of heparin [Lindahl, Bäckström, Thunberg & Leder (1980) Proc. Natl. Acad. Sci. U.S.A. 77, 6551-6555], could be demonstrated. A significantly lower proportion of the chains derived from transformed cells bound with high affinity to antithrombin, and a corresponding decrease in the amount of incorporated 3-O-sulphate was observed.

Topics: Animals; Antithrombins; Basement Membrane; Cell Line; Cell Transformation, Neoplastic; Chromatography, Affinity; Chromatography, Gel; Chromatography, High Pressure Liquid; Epithelium; Female; Glycosaminoglycans; Heparitin Sulfate; Mammary Glands, Animal; Mice; Nitrous Acid; Oligosaccharides; Proteoglycans

Treatment of normal human (WI-38) cells with exogenous heparan sulfate (HS) reduced cell growth and incorporation of radio-isotope-labeled thymidine (TdR) into DNA. In spite that growth of their transformants (WI-38 CT-1) was enhanced by HS treatment, transformed cells also decreased in TdR incorporation thereby. This peculiar observation was explained by a reduction of TdR uptake, leading to a decrease in specific radioactivity of newly synthesized DNA. The changes in cell growth and TdR incorporation by HS treatment were revealed to be similar to the changes with increasing cell density rather than by serum starvation.

Topics: Biological Transport; Cell Division; Cell Transformation, Neoplastic; Cells, Cultured; Contact Inhibition; DNA; Fibroblasts; Glycosaminoglycans; Heparitin Sulfate; Humans; Hyaluronic Acid; Thymidine

The distribution of basement membrane (BM) markers, type IV collagen, laminin (LM), heparan sulphate proteoglycan (HSP) and fibronectin (FN) has been studied by indirect immunofluorescence using specific antibodies, in tumoural pathology. The disrupted pattern of BM by these markers in severe dysplastic lesions of the breasts, the bronchi and uterine cervix provides evidence for malignancy. In invasive carcinomas, there is generally a loss of these BM components, with FN persisting in the stroma. The loss of these markers in BM is concomitant and superimposable in double staining studies. In embryonic tumours, the presence of BM markers is related to a mesenchymal differentiation of malignant cells with pericellular FN and/or maturation towards organoid structures with BM. In sarcomas, there is a loss of the pericellular BM staining around most transformed muscular and Schwann cells and adipocytes. The persistence of this labelling in some well-differentiated areas can help to diagnose the nature of the sarcoma. The persistence of intercellular filaments of FN corresponds to the mesenchymal and/or sarcomatous nature of undifferentiated anaplastic proliferations.

Topics: Basement Membrane; Breast Neoplasms; Cell Transformation, Neoplastic; Collagen; Female; Fibronectins; Fluorescent Antibody Technique; Heparitin Sulfate; Humans; Laminin; Lung Neoplasms; Male; Neoplasms; Neoplasms, Germ Cell and Embryonal; Sarcoma

Protamine sulfate reversibly inhibits serum-induced mitogenic stimulation of several nontransformed and neoplastic cell types in vitro. Fifty percent inhibition was induced by approximately 120-150 micrograms protamine sulfate/ml. Cells were affected directly, and inhibition depended on the duration of cell exposure. Heparin, chondroitin sulfate, heparan sulfate, and dextran sulfate neutralized protamine sulfate effects during the early stages of treatment. Nontransformed cells [bovine aortic endothelial cells, adult human gingival fibroblasts (strains 423 and 1101), fetal rat skin (strain 921-K) and muscle fibroblasts] required longer exposure to induce inhibition than did neoplastic cells [rat 3-methylcholanthrene-induced fibrosarcoma cell lines (MCA-6 and MCA-9), a macrophage-like cell line (NCTC-3749), Walker 256 rat carcinoma cells (ATCC-CCL-38), rat Morris hepatoma cells (ATCC-CCL-144), murine melanoma cells (B16), and rat bladder squamous cell carcinoma cells (804-G)]. Other polycationic compounds, including histone type VIII-S, poly-L-lysine, poly-L-arginine, and protamine (free base), were also effective inhibitors, whereas the basic proteins cytochrome c and lysozyme had no effect. Poly-L-histidine, poly-L-glutamic acid, poly-L-aspartic acid, and dextran blue also had no inhibitory effect.

Topics: Animals; Cell Division; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Chondroitin Sulfates; Culture Media; Dextran Sulfate; Dextrans; DNA Replication; Heparin; Heparitin Sulfate; Humans; Kinetics; Mice; Neoplasms, Experimental; Peptides; Protamines; Rats

A rat hepatoma cell line was shown to synthesize heparan sulfate and chondroitin sulfate proteoglycans. Unlike cultured hepatocytes, the hepatoma cells did not deposit these proteoglycans into an extracellular matrix, and most of the newly synthesized heparan sulfate proteoglycans were secreted into the culture medium. Heparan sulfate proteoglycans were also found associated with the cell surface. These proteoglycans could be solubilized by mild trypsin or detergent treatment of the cells but could not be displaced from the cells by incubation with heparin. The detergent-solubilized heparan sulfate proteoglycan had a hydrophobic segment that enabled it to bind to octyl-Sepharose. This segment could conceivably anchor the molecule in the lipid interior of the plasma membrane. The size of the hepatoma heparan sulfate proteoglycans was similar to that of proteoglycans isolated from rat liver microsomes or from primary cultures of rat hepatocytes. Ion-exchange chromatography on DEAE-Sephacel indicated that the hepatoma heparan sulfate proteoglycans had a lower average charge density than the rat liver heparan sulfate proteoglycans. The lower charge density of the hepatoma heparan sulfate can be largely attributed to a reduced number of N-sulfated glucosamine units in the polysaccharide chain compared with that of rat liver heparan sulfate. Hepatoma heparan sulfate proteoglycans purified from the culture medium had a considerably lower affinity for fibronectin-Sepharose compared with that of rat liver heparan sulfate proteoglycans. Furthermore, the hepatoma proteoglycan did not bind to the neoplastic cells, whereas heparan sulfate from normal rat liver bound to the hepatoma cells in a time-dependent reaction. The possible consequences of the reduced sulfation of the heparan sulfate proteoglycan produced by the hepatoma cells are discussed in terms of the postulated roles of heparan sulfate in the regulation of cell growth and extracellular matrix formation.

Topics: Animals; Cell Line; Cell Transformation, Neoplastic; Fibronectins; Glycosaminoglycans; Heparitin Sulfate; Liver Neoplasms, Experimental; Microsomes; Proteoglycans; Rats

Topics: Animals; Cell Line; Cell Transformation, Neoplastic; Glycosaminoglycans; Heparitin Sulfate; Mice; Sulfurtransferases; Trypsin

Previous studies showed that cell surface heparan sulfate (HS) is involved in density-dependent growth regulation of normal human cells. In this study the effects of HS on proliferation of transformed cells were studied in vitro. Exogenously added HS prepared from normal tissues (rat kidney and bovine kidney) enhanced growth of transformed human and animal cells (gamma ray- or virus-transformed WI-38, and HeLa cells and chemically induced mouse hepatoma cells), but inhibited that of normal human and animal cells (WI-38, 3T3, and rabbit liver cells). HS was less effective on growth of both normal and transformed human cells at higher cell density. Although the exogenous HS did not bind to cells tightly, HS was found to affect cell growth not by modulation of growth-related substances in the medium, but through contact with the cell surface. HS preparation from tumour cells (mouse hepatoma cells) exerted similar effects on cell growth. Heparin, structurally similar to HS, inhibited growth of both normal and transformed human cells. These findings suggest that: 1. HS plays a particular function in contact regulation of cell proliferation. 2. Transformation-related changes in the structure of HS molecules do not much affect the function of HS. 3. The cellular transformation, however, is accompanied by alteration in the growth-regulating system sensitive to extracellular HS.

Topics: Animals; Cell Division; Cell Transformation, Neoplastic; Cells, Cultured; Glycosaminoglycans; Heparin; Heparitin Sulfate; Humans; Mice; Rabbits; Thymidine

This review describes some structural details and the metabolism of the sulfated glycosaminoglycans in animal cells in a variety of physiological conditions and presents views on the possible role that these compounds may play in cell self-recognition, neoplastic transformation and in the control of cell growth.

Topics: Animals; Cell Adhesion; Cell Division; Cell Physiological Phenomena; Cell Transformation, Neoplastic; Cells, Cultured; Chemical Phenomena; Chemistry; Chondroitin Sulfates; Dermatan Sulfate; Glycosaminoglycans; Heparitin Sulfate

Proteoglycans deposited in the basal lamina of [14C] glucosamine-labeled normal and [3H]glucosamine-labeled transformed mouse mammary epithelial cells grown on type I-collagen gels, were extracted in 4 M guanidinium chloride and cofractionated over Sepharose CL 4B. The heparan sulfate chains carried by these proteoglycans were isolated by treatment with alkaline borohydride, protease K, chondroitinase ABC, and cetylpyridinium chloride precipitation. Heparan sulfate isolated from transformed cell cultures consistently eluted from DEAE-cellulose at lower salt concentrations and was of smaller apparent Mr when chromatographed over Sepharose CL 6B, than heparan sulfate of normal cell cultures. Experiments using doubly labeled cultures ([3H]glucosamine and [35S]sulfate) demonstrated an approximately 30% reduction in the sulfate/hexosamine ratio in heparan sulfate derived from transformed cultures. Both N- and O-sulfate were decreased. The decreased Mr and decreased sulfation of heparan sulfate upon transformation appear sufficient to explain the altered heparan sulfate/chondroitin sulfate ratios previously observed in these cells. These changes may have implications for the molecular interactions in which these proteoglycans are normally engaged during basal lamina assembly, and cause the poor basal lamina formation displayed by these transformed cells.

Topics: Animals; Basement Membrane; Cell Line; Cell Transformation, Neoplastic; Epithelium; Female; Glucosamine; Glycosaminoglycans; Heparitin Sulfate; Mammary Glands, Animal; Mice; Proteoglycans; Tritium

Topics: Adult; Animals; Cell Division; Cell Transformation, Neoplastic; Cell Transformation, Viral; Cells, Cultured; Chemical Phenomena; Chemistry; Chondroitin Sulfates; Dogs; Glycosaminoglycans; Guinea Pigs; Heparitin Sulfate; Humans; Liver; Mice; Rabbits; Regeneration; Swine; Tissue Distribution

Albumin-producing rat liver parenchymal cell clones (BB and BC) and their subclones in the confluent culture synthesized heparan sulfate as the major component and dermatan sulfate, chondroitin sulfate and hyaluronic acid as the minor ones. Their relative contents were similar to those present in the rat liver. Analyses of glycosaminoglycans synthesized by subclone cells (BB1S) at various cell densities, cell growth phases and passage levels have shown that relative content of heparan sulfate remained constant, suggesting that the epithelial cell possesses a stable heparan sulfate-producing capacity. On the other hand, the level of hyaluronic acid production was high at low cell density, though it remained constant during cell proliferation. Chemically transformed rat liver parenchymal cells (M) produced relatively higher amount of chondroitin sulfate than non-transformed cells did, as observed with 4-nitroquinoline-1-oxide-transformed 3T3 cells, compared to 3T3 714 cells. The results obtained on this study strongly suggest that the liver parenchymal cells synthesize a major part of the glycosaminoglycans of the liver and chondroitin sulfate production is closely related to cellular proliferations.

Topics: Animals; Cell Division; Cell Transformation, Neoplastic; Chondroitin Sulfates; Clone Cells; Dermatan Sulfate; Glycosaminoglycans; Heparitin Sulfate; Hyaluronic Acid; Liver; Rats

Topics: Cell Line; Cell Transformation, Neoplastic; Chondroitin; Chondroitin Sulfates; Disaccharides; Glycosaminoglycans; Heparitin Sulfate

Footpad adhesion sites pinch off from the rest of the cell surface during EGTA-mediated detachment of normal or virus-transformed murine cells from their tissue culture substrates. In these studies, highly purified trypsin and testicullar hyaluronidase were used to investigate the selective destruction or solubilization of proteins and polysaccharides in this substrate-attached material (SAM). Trypsin-mediated detachment of cells or trypsinization of SAM after EGTA-mediated detachment of cells resulted in the following changes in SAM composition: (a) solubilization of 50-70% of the glycosaminoglycan polysaccharide with loss of only a small fraction of the protein, (b) selective loss of one species of glycosaminoglycan-associated protein in longterm radiolabeled preparations, (c) no selective loss of the LETS glycoprotein or cytoskeletal proteins in longterm radiolabeled preparations, and (d) selective loss of one species of glycosaminoglycan-associated protein, a protion of the LETS glycoprotein, and proteins Cd (mol wt 47,000 and Ce' (mol wt 39,000) in short term radiolabeled preparations. Digestion of SAM with testicular hyaluronidase resulted in: (a) almost complete solubilization of the hyaluronate and chondroitin sulfate moieties from long term radiolabeled SAM with minimal loss of heparan sulfate, (b) solubilization of a small portion of the LETS glycoprotein and the cytoskeletal proteins from longterm radiolabeled SAM, (c) resistance to solubilization of protein and polysaccharide in reattaching cell SAM which contains principally heparan sulfate, and (d) complete solubilization of the LETS glycoprotein in short term radiolabeled preparations with no loss of cytoskeletal proteins. Thus, there appear to be two distinct pools of LETS in SAM, one associated in some unknown fashion with hyaluronate-chondroitin sulfate complexes, and a second associated with some other component in SAM, perhaps heparan sulfate. These data, together with other results, suggest that the cell-substrate adhesion process may be mediated principally by a heparan sulfate--LETS complex and that hyaluronate-chondroitin sulfate complexes may be important in the detachability of cells from the serum-coated substrate by destabilizing LETS matrices at posterior footpad adhesion sites.

Topics: Animals; Cell Adhesion; Cell Line; Cell Separation; Cell Transformation, Neoplastic; Cell Transformation, Viral; Chondroitin Sulfates; Glycoproteins; Glycosaminoglycans; Heparitin Sulfate; Hyaluronic Acid; Mice; Neoplasm Proteins; Solubility

Heparan sulfate from the surface of a variety of mouse cells at different cell densities was examined by ion-exchange chromatography. The results of this analysis show that: (1) The heparan sulfate from new isolates of Swiss 3T3 cells transformed by SV40 virus (a DNA tumor virus) elutes from DEAE-cellulose at a lower ionic strength than that from the parent cell type. This finding confirms our earlier observation with an established SV40-transformed cell line (Underhill and Keller, '75) and eliminates the possibility that this change is caused by extended passage in culture. (2) For both parent and transformed 3T3 cells, the heparan sulfates from low and high density cultures were the same as judged by chromatography on DEAE-cellulose. This result demonstrates that the transformation-dependent change which we have observed is independent of cell density. (3) The heparan sulfate from Balb/c 3T3 cells transformed with Kirsten murine sarcoma virus (an RNA tumor virus) elutes from DEAE-cellulose prior to that from parent Balb/c 3T3 cells. This result extends the transformation dependent change in heparan sulfate to the Balb/c 3T3 cell line and to cells transformed with an RNA virus.

Topics: Cell Count; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Chromatography, Ion Exchange; Glycosaminoglycans; Heparitin Sulfate

Glycosaminoglycans (GAG's) were released by trypsin from the surface of cultured mouse cells (3T3) in two different growing states: during log-growth phase and during resting due to serum starvation. Doubly labelled molecules from resting cells were compared with those from growing as well as from trnsformed cells. Reproducible differences in the elution pattern during ion exchange chromatography and in susceptibility to specific hydrolytic enzymes have been demonstrated: the GAGs pattern of growing normal cells is similar to the pattern of the cells transformed by either Polyoma or SV-40 viruses and very different from the pattern of resting cells. Growing and transformed 3T3 show a relatively low amount of trypsin removable heparan sulphate (HS) and a relatively high amount of hyaluronic acid (HA) while resting cells exhibit an opposite ratio between the two GAG'S. The lowering of HS and the increase of HA in the cell coat is therefore suspected to be more dependent upon growth than upon transformation.

Topics: Animals; Cell Division; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Chondroitin Sulfates; Glycopeptides; Glycosaminoglycans; Heparitin Sulfate; Hyaluronic Acid; Mice; Polyomavirus; Simian virus 40; Trypsin

The relative amount of sulfated glycosaminoglycans associated with the cell layer of parent and SV40-transformed Swiss mouse 3T3 cells was determined from the incorporation of labeled sulfate (35SO4) into macromolecular material. In cultures of SV40-transformed cells, the glycosaminoglycan content per cell was constant over a wide range of densities. In cultures of parent 3T3 cells, the glycosaminoglycan content per cell increased directly with density, the highest values being found in contact-inhibited cultures. At high cell densities, the glycosaminoglycan content of 3T3 cells was several-fold higher than that for SV40-transformed cells. Most of the density-dependent increase in glycosaminoglycans of 3T3 cells was accounted for by chondroitin sulfate (dermatan sulfate) which was over 6-fold higher in confluent cultures than in low density cultures.

Topics: Cell Count; Cell Line; Cell Transformation, Neoplastic; Chondroitin; Chondroitin Sulfates; Glycosaminoglycans; Heparitin Sulfate

Topics: Animals; Cell Differentiation; Cell Division; Cell Membrane; Cell Transformation, Neoplastic; Fibroblasts; Glycosaminoglycans; Heparitin Sulfate; Membrane Lipids; Membrane Proteins; Mitosis; Models, Biological; Nucleotides, Cyclic; Transcription, Genetic; Virus Replication

Mouse 3T3 cells and their Simian Virus 40-transformed derivatives (3T3SV) were used to assess the relationship of transfromation, cell density, and growth control to the cellular distribution of newly synthesized glycosaminoglycan (GAG). Glucosamine- and galactosamine-containing GAG were labeled equivalently by [3H=A1-glucose regardless of culture type, allowing incorporation into the various GAG to be compared under all conditions studied. Three components of each culture type were examined: the cells, which contain the bulk of newly synthesized GAG and are enriched in chondroitin sulfate and heparan sulfate; cell surface materials released by trypsin, which contain predominantly hyaluronic acid; and the media , which contain predominantly hyaluronic acid and undersulfated chondroitin sulfate. Increased cell density and viral transformation reduce incorporation into GAG relative to the incorporation into other polysaccharides. Transformation, however, does not substantially alter the type or distribution of newly synthesized GAG; the relative amounts and cellular distributions were very similar in 3T3 and 3T3SV cultures growing at similar rates at low densities. On the other hand, increased cell density as well as density-dependent growth inhibition modified the type and distribution of newly synthesized GAG. At high cell densities both cell types showed reduced incorporation into hyaluronate and an increase in cellular GAG due to enhanced labeling of chondroitin sulfate and heparan sulfate. These changes were more marked in confluent 3T3 cultures which also differed in showing substantially more GAG label in the medium and in chondroitin-6-sulfate and heparan sulfate at the cell surface. Since cell density and possibly density-dependent inhibition of growth but not viral transformation are major factors controlling the cellular distribution and type of newly synthesized GAG, differences due to GAG's in the culture behavior of normal and transformed cells may occur only at high cell density. The density-induced GAG alterations most likely involved are increased condroitin-6-sulfate and heparan sulfate and decreased hyaluronic acid at the cell surface.

Topics: Cell Count; Cell Division; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Chondroitin Sulfates; Contact Inhibition; Culture Media; Glucose; Glycosaminoglycans; Heparitin Sulfate; Hyaluronic Acid

Topics: Cell Adhesion; Cell Differentiation; Cell Division; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Glycosaminoglycans; Heparin; Heparitin Sulfate; Hyaluronic Acid; Models, Biological

Topics: Cell Division; Cell Line; Cell Transformation, Neoplastic; Chondroitin; Chondroitin Sulfates; Galactosamine; Glucosamine; Glycopeptides; Glycosaminoglycans; Heparitin Sulfate; Hyaluronic Acid

Topics: Animals; Cell Transformation, Neoplastic; Cells, Cultured; Chondroitin; Chromatography, DEAE-Cellulose; Chromatography, Gel; Glycosaminoglycans; Heparitin Sulfate; Hexosamines; Hyaluronic Acid; Mice; RNA; Simian virus 40; Trypsin; Uridine

The rate of hyaluronic acid and sulfated mucopolysaccharide production was measured for hamster embryo fibroblasts and for general oncogenic lines derived by virus transformation. A striking increase in both the rate of hyaluronic acid synthesis and the amount of cell-associated polymer was observed after transformation by herpes simplex type-2 or SV40 virus. Although no corresponding change was observed for the sulfated polysaccharides, the proportion of heparan sulfate increased significantly after transformation.

Topics: Animals; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Chondroitin; Cricetinae; Embryo, Mammalian; Fibroblasts; Glucosamine; Glycosaminoglycans; Haplorhini; Heparin; Heparitin Sulfate; Hexosaminidases; Hyaluronic Acid; Hyaluronoglucosaminidase; Kidney; Kinetics; Simian virus 40; Simplexvirus; Tritium