heparitin-sulfate and Adenocarcinoma

Topics: Adenocarcinoma; Animals; Basement Membrane; Collagen; Glucuronidase; Glycosaminoglycans; Glycoside Hydrolases; Heparitin Sulfate; Humans; Mammary Neoplasms, Experimental; Melanoma; Neoplasm Metastasis; Rats

Prostate cancer is impacting many men globally. It is a disease that has no effective treatment is available in the market. The understanding of the biophysical and biochemical aspects of the disease and the mechanism that allow it to metastasize is key to finding an effect treatment. Maintenance or pretreatment drug as well as a post treatment drug can be effective to avoid or delay the disease from appearing. The polysaccharides and monosaccharides polymers combined with vitamins can be the ingredient to developing the treatment. There are many evidences that investigators examined the individual components of the therapy proposed but never a combination of all these therapies. The one item that is not discussed is how to formulate the ingredient into an effective form which is a proprietary work being conducted currently. Nevertheless, the hypothesis seems reasonable to us and worth sharing with the scientific community.

Topics: Adenocarcinoma; Amygdalin; Antineoplastic Combined Chemotherapy Protocols; Cell Division; Drug Combinations; Drug Synergism; Extracellular Matrix; Glycosaminoglycans; Heparitin Sulfate; Humans; Injections, Intravenous; Male; Models, Biological; Myocytes, Smooth Muscle; Polysaccharides; Prostate; Prostatic Neoplasms; Vitamin A; Vitamin E

Glycans play a critical role in physiological and pathological processes through interaction with a variety of ligands. Altered expression and dysregulation of these molecules can cause aberrant cellular function such as malignancy. Glycomics provide information of the structure and function of glycans, glycolipids, and glycoproteins such as proteoglycans, and may help to predict cancer development and progression as biomarkers. In this report, we compared the expression of proteoglycans, the content and structure of glycosaminoglycans and glycolipids between patient-matched normal and cancer tissues obtained from colon cancer patients. Tumor-related proteoglycans, glypican-3, and syndecan-1 showed downregulation in cancer tissues compared to normal tissues. In cancer tissue, the total amount of chondroitin sulfate (CS)/dermatan sulfate and heparan sulfate were lower and, interestingly, the level of disaccharide units of both 4S6S (CS-E) and 6S (CS-C) were higher compared to normal tissue. Also, overall lipids including glycolipids, a major glycomics target, were analyzed by hydrophilic interaction liquid chromatography mass spectrometry. Increase of lyso-phosphatidylcholine (phospholipid), sphingomyelin (sphigolipid), and four types of glycolipids (glucosylceramide, lactosylceramide, monosialic acid ganglioside, and globoside 4) in cancer tissue showed the possibility as potential biomarkers in colon cancer. While requiring the need for careful interpretation, this type of broad investigation gives us a better understanding of pathophysiological roles on glycosaminoglycans and glycolipids and might be a powerful tool for colon cancer diagnosis.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Antigens, CD; Biomarkers; Carbohydrate Sequence; Case-Control Studies; Chondroitin Sulfates; Colonic Neoplasms; Dermatan Sulfate; Female; Gangliosides; Gene Expression Regulation, Neoplastic; Globosides; Glucosylceramides; Glypicans; Heparitin Sulfate; Humans; Lactosylceramides; Lysophosphatidylcholines; Male; Middle Aged; Molecular Sequence Data; Sphingomyelins; Syndecan-1

To determine the expression pattern and prognostic value of heparan sulfate in gastric cancer.. The 10E4 antiheparan sulfate monoclonal antibody was used to examine the expression pattern of heparan sulfate in tissue microarrays consisting of 162 cases of gastric carcinoma by immunohistochemistry. The immunoreactivities of both epithelial and stromal components of the specimens were examined and analysed statistically for significant associations with clinicopathological parameters, including histological grade of the tumour, extent of cancer infiltration and presence of lymph-node metastases, lymphovascular invasion, perineural invasion, perforation of gastric wall and stromal reaction. The potential use of heparan sulfate as a predictive factor for patient survival was also evaluated.. Reduced expression of heparan sulfate in the epithelial component was associated with higher histological grades of gastric cancer as well as the presence of more extensive tumour infiltration. Furthermore, this decrease in heparan sulfate expression was found to be predictive of reduced patient survival after tumour recurrence.. The data suggest that heparan sulfate may play an important role in regulating the biology of gastric cancer, and that it may be a useful prognostic marker of this tumour.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Cell Differentiation; Epidemiologic Methods; Female; Heparitin Sulfate; Humans; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Invasiveness; Prognosis; Stomach Neoplasms; Tumor Cells, Cultured

The epidermal growth factor receptor (EGFR) mutation status is a validated biomarker for the stratification of EGFR-tyrosine kinase inhibitor (EGFR-TKIs) treatment in patients with non-small cell lung cancer (NSCLC); however, its use is limited in patients with wild-type EGFR, and new biomarkers are needed. We hypothesized that the serum concentration of heparan sulfate (HS), which activates oncogenic growth factor receptor signaling through EGFR and non-EGFR signaling pathways, may be a novel glycobiological biomarker for EGFR-TKIs treatment in NSCLC.. The pretreatment serum HS concentrations were determined using enzyme-linked immunosorbent assay in 83 patients with stage IV non-small cell lung adenocarcinoma who received EGFR-TKIs treatment. The relationship between the serum HS concentrations and patient characteristics, tumor response, progression-free survival (PFS), and overall survival (OS) were analyzed.. Patient sex, performance status, smoking history, and EGFR mutation status were associated with tumor response. The serum HS concentrations were significantly higher among patients with progressive disease than among those without progressive disease (p = 0.003). Furthermore, the serum HS concentrations were strongly associated with a poor PFS and OS in a univariate Cox analysis (p = 0.0022 and p = 0.0003, respectively). A stratified multivariate Cox model according to the EGFR mutation status showed that higher HS concentrations were significantly associated with a shorter PFS and OS (p = 0.0012 and p = 0.0003).. We concluded that a high-serum HS concentration was strongly related to a poor treatment outcome of EGFR-TKIs and may be a promising noninvasive and repeatable glycobiological biomarker in cancer treatment.

Topics: Adenocarcinoma; Aged; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; ErbB Receptors; Erlotinib Hydrochloride; Female; Gefitinib; Heparitin Sulfate; Humans; Lung Neoplasms; Male; Mutation; Neoplasm Staging; Protein Kinase Inhibitors; Quinazolines; Retrospective Studies; Treatment Failure

Hedgehog (Hh) proteins are morphogens that mediate many developmental processes. Hh signaling is significant for many aspects of embryonic development, whereas dysregulation of this pathway is associated with several types of cancer. Hh proteins require heparan sulfate proteoglycans (HSPGs) for their normal distribution and signaling activity. Here, we have used molecular modeling to examine the heparin-binding domain of sonic hedgehog (Shh). In biochemical and cell biological assays, the importance of specific residues of the putative heparin-binding domain for signaling was assessed. It was determined that key residues in human (h) Shh involved in heparin and HSPG syndecan-4 binding and biological activity included the well known cationic Cardin-Weintraub motif (lysines 32-38) but also a previously unidentified major role for lysine 178. The activity of Shh mutated in these residues was tested by quantitation of alkaline phosphatase activity in C3H10T1/2 cells differentiating into osteoblasts and hShh-inducible gene expression in PANC1 human pancreatic ductal adenocarcinoma cells. Mutated hShhs such as K37S/K38S, K178S, and particularly K37S/K38S/K178S that could not interact with heparin efficiently had reduced signaling activity compared with wild type hShh or a control mutation (K74S). In addition, the mutant hShh proteins supported reduced proliferation and invasion of PANC1 cells compared with control hShh proteins, following endogenous hShh depletion by RNAi knockdown. The data correlated with reduced Shh multimerization where the Lys-37/38 and/or Lys-178 mutations were examined. These studies provide a new insight into the functional roles of hShh interactions with HSPGs, which may allow targeting this aspect of hShh biology in, for example, pancreatic ductal adenocarcinoma.

Topics: Adenocarcinoma; Amino Acid Motifs; Amino Acid Substitution; Cell Line, Tumor; Hedgehog Proteins; Heparitin Sulfate; Humans; Mutation, Missense; Osteoblasts; Pancreatic Neoplasms; Protein Multimerization; Protein Structure, Tertiary; Signal Transduction; Syndecan-4

Heparin/heparan sulfate (HS) proteoglycans are found in the extracellular matrix (ECM) and on the cell surface. A considerable body of evidence has established that heparin and heparan sulfate proteoglycans (HSPGs) interact with numerous protein ligands including fibroblast growth factors, vascular endothelial growth factor (VEGF), cytokines, and chemokines. These interactions are highly dependent upon the pattern of sulfation modifications within the glycosaminoglycan chains. We previously cloned a cDNA encoding a novel human endosulfatase, HSulf-2, which removes 6-O-sulfate groups on glucosamine from subregions of intact heparin. Here, we have employed both recombinant HSulf-2 and the native enzyme from conditioned medium of the MCF-7-breast carcinoma cell line. To determine whether HSulf-2 modulates the interactions between heparin-binding factors and heparin, we developed an ELISA, in which soluble factors were allowed to bind to immobilized heparin.. Our results show that the binding of VEGF, FGF-1, and certain chemokines (SDF-1 and SLC) to immobilized heparin was abolished or greatly diminished by pre-treating the heparin with HSulf-2. Furthermore, HSulf-2 released these soluble proteins from their association with heparin. Native Sulf-2 from MCF-7 cells reproduced all of these activities.. Our results validate Sulf-2 as a new tool for deciphering the sulfation requirements in the interaction of protein ligands with heparin/HSPGs and expand the range of potential biological activities of this enzyme.

Topics: Adenocarcinoma; Breast Neoplasms; Cell Line, Tumor; Chemokine CCL21; Chemokine CXCL12; Chemokines, CC; Chemokines, CXC; Culture Media, Conditioned; DNA, Complementary; Enzyme-Linked Immunosorbent Assay; Female; Fibroblast Growth Factor 1; Fibroblast Growth Factor 2; Heparin; Heparitin Sulfate; Humans; Interleukin-8; Neoplasm Proteins; Protein Binding; Recombinant Fusion Proteins; Serum Albumin, Bovine; Sulfatases; Sulfotransferases; Vascular Endothelial Growth Factor A

Proliferation of fibroblasts (desmoplastic reaction) in the lung adenocarcinomas is an important phenomenon that correlates with metastases and poor prognosis. Because basement membranes are often involved in the desmoplastic areas and many cytokines have binding capacity to basement membrane molecules, we hypothesized that basement membrane modify the paracrine effects between cancer cells and fibroblasts via the fibrogenic cytokines and this hypothesis was experimentally investigated.. The effects of conditioned media derived from ten lung carcinoma cell lines and normal airway epithelial cells on DNA synthesis of fetal lung fibroblasts were determined. We focused on fibroblast growth factor 2 (FGF-2) as the candidate paracrine cytokines and examined their diffusion through an experimental basement membrane matrix model, Matrigel.. All the conditioned media promoted DNA synthesis of fetal lung fibroblasts. Detection by ELISA methods and the neutralizing antibodies suggested that FGF-2 was one of the responsible factors for the growth promotion. Diffusion of FGF-2 across the polycarbonate membrane was suppressed by coating with Matrigel. When FGF-2-secreting A549 cells were covered with Matrigel, FGF-2 was stored in Matrigel and its diffusion into the culture media was significantly reduced. Binding of FGF-2 to Matrigel was completely blocked by a basic protein, protamine sulfate. In the presence of protamine sulfate in Matrigel overlaid on A549 cells, diffusion of FGF-2 increased 7-fold as much as that without overlaid Matrigel.. These results suggest that the basement membrane acts as a barrier to the diffusion and a reservoir of cytokines secreted by cancer cells, and that the subsequent degradation of the basement membrane by cancer cells could release the stored cytokines and promote growth of fibroblasts.

Topics: Adenocarcinoma; Antibodies, Blocking; Basement Membrane; Cell Division; Cell Line, Tumor; Collagen; Culture Media, Conditioned; DNA; DNA Replication; Dose-Response Relationship, Drug; Drug Combinations; Fibroblast Growth Factor 2; Fibroblasts; Heparitin Sulfate; Humans; Interleukin-1; Laminin; Lung; Lung Neoplasms; Paracrine Communication; Protamines; Proteoglycans; Recombinant Proteins

Heparanase (HPSE-1) is involved in the degradation of both cell-surface and extracellular matrix (ECM) heparan sulfate (HS) in normal and neoplastic tissues. Degradation of heparan sulfate proteoglycans (HSPG) in mammalian cells is dependent upon the enzymatic activity of HPSE-1, an endo-beta-d-glucuronidase, which cleaves HS using a specific endoglycosidic hydrolysis rather than an eliminase type of action. Elevated HPSE-1 levels are associated with metastatic cancers, directly implicating HPSE-1 in tumor progression. The mechanism of HPSE-1 action to promote tumor progression may involve multiple substrates because HS is present on both cell-surface and ECM proteoglycans. However, the specific targets of HPSE-1 action are not known. Of particular interest is the relationship between HPSE-1 and HSPG, known for their involvement in tumor progression. Syndecan-1, an HSPG, is ubiquitously expressed at the cell surface, and its role in cancer progression may depend upon its degradation. Conversely, another HSPG, perlecan, is an important component of basement membranes and ECM, which can promote invasive behavior. Down-regulation of perlecan expression suppresses the invasive behavior of neoplastic cells in vitro and inhibits tumor growth and angiogenesis in vivo. In this work we demonstrate the following. 1) HPSE-1 cleaves HS present on the cell surface of metastatic melanoma cells. 2) HPSE-1 specifically degrades HS chains of purified syndecan-1 or perlecan HS. 3) Syndecan-1 does not directly inhibit HPSE-1 enzymatic activity. 4) The presence of exogenous syndecan-1 inhibits HPSE-1-mediated invasive behavior of melanoma cells by in vitro chemoinvasion assays. 5) Inhibition of HPSE-1-induced invasion requires syndecan-1 HS chains. These results demonstrate that cell-surface syndecan-1 and ECM perlecan are degradative targets of HPSE-1, and syndecan-1 regulates HPSE-1 biological activity. This suggest that expression of syndecan-1 on the melanoma cell surface and its degradation by HPSE-1 are important determinants in the control of tumor cell invasion and metastasis.

Topics: Adenocarcinoma; Animals; Cell Membrane; Colorectal Neoplasms; Extracellular Matrix; Glucuronidase; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Hydrogen-Ion Concentration; Melanoma; Membrane Glycoproteins; Mice; Neoplasm Invasiveness; Neoplasm Metastasis; Proteoglycans; Recombinant Proteins; Substrate Specificity; Syndecan-1; Syndecans; Tumor Cells, Cultured

Topics: Adenocarcinoma; Angiogenesis Inducing Agents; Basement Membrane; Breast Neoplasms; Disease Progression; Enzyme Inhibitors; Female; Glucuronidase; Growth Substances; Heparitin Sulfate; Humans; Neoplasm Invasiveness; Neoplasm Metastasis; RNA, Catalytic; Transfection

The tyrosine kinase receptor c-Met and its ligand HGF/SF, ezrin, and splice variants of CD44 have independently been identified as tumor metastasis-associated proteins. We now show that these proteins cooperate. A CD44 isoform containing variant exon v6 sequences is strictly required for c-Met activation by HGF/SF in rat and human carcinoma cells, in established cell lines as well as in primary keratinocytes. CD44v6-deficient tumor cells were unable to activate c-Met unless they were transfected with a CD44v6-bearing isoform. Antibodies to two v6-encoded epitopes inhibited autophosphorylation of c-Met by interfering with the formation of a complex formed by c-Met, CD44v6, and HGF/SF. In addition, signal transduction from activated c-Met to MEK and Erk required the presence of the cytoplasmic tail of CD44 including a binding motif for ERM proteins. This suggests a role for ERM proteins and possibly their link to the cortical actin cytoskeleton in signal transfer.

Topics: Adenocarcinoma; Cell Line; Colonic Neoplasms; Heparitin Sulfate; Hepatocyte Growth Factor; Humans; Hyaluronan Receptors; Keratinocytes; Protein Isoforms; Proto-Oncogene Proteins c-met; Signal Transduction

The paper shows the ability of the fluorochrome tris(2,2'-bipyridine) ruthenium (II) (Rubipy) to detect heparan sulfate, heparin, and heparinase activity of M3 murine mammary adenocarcinoma cells as well as bacterial heparinases I, II, and III in native polyacrylamide gel electrophoresis (PAGE). The technique is based on the electrophoretic mobility of high molecular weight heparins and subsequent staining with Rubipy (50 micrograms/mL). The minimum content of heparin detected by fluorescence in a UV transilluminator was 25-50 ng. The number of Rubipy molecules bound to heparin, determined in relationship to the number of disaccharide units (DU), showed that two to six heparin disaccharide units are bound by each fluorochrome molecule. Scatchard plot analysis showed one Rubipy-binding site (Kd = (8.56 +/- 2.97) x 10(-5) M). Heparinase activity was determined by densitometric analysis of the fluorescence intensity of the heparin-containing band of the gel. While heparinase I (EC 4.2.2.7.) degraded heparin and, to a lower degree, partially N-desulfated N-acetylated heparin (N-des N-Ac), heparinase II (no EC number) could efficiently degrade heparan sulfate (HS) and partially N-des N-Ac heparin. Finally, heparinase III (EC 4.2.2.8.) degraded HS almost exclusively. Only heparin and N-des N-Ac heparin were substrates for M3 tumor cell heparinases. We describe a qualitative, sensitive and simple method to detect heparinase activity and determine its substrate specificity using Rubipy fluorescence with heparin and heparan sulfate in multiple biological samples tested in parallel.

Topics: 2,2'-Dipyridyl; Adenocarcinoma; Animals; Coordination Complexes; Electrophoresis, Polyacrylamide Gel; Female; Fluorescent Dyes; Heparin; Heparin Lyase; Heparitin Sulfate; Mammary Neoplasms, Animal; Mice; Molecular Structure; Polysaccharide-Lyases; Tumor Cells, Cultured

Syndecan-1 binds basic fibroblast growth factor (bFGF), modulates neovascularization, plays a role in epithelial differentiation and is up-regulated by WT1. Malignant mesothelioma of the pleura is one of the most aggressive tumours known and expresses high levels of angiogenic growth factors. This study has analysed syndecan-1 expression in mesothelioma tumours and cell lines by immunohistochemistry and immunoblotting, using anti-syndecan-1 antibody directed against the core protein, and has examined its relation to morphology, bFGF, WT1, and intra-tumoural microvascular density (IMD). Shedding of syndecan-1 in the conditioned medium of mesothelioma cell lines was detected in variable amounts. These studies indicate that (1) there is no correlation of syndecan-1 with either bFGF expression or IMD in mesotheliomas in vivo; (2) syndecan-1 is strongly expressed in the epithelial type of mesothelioma and in the epithelial component of biphasic mesotheliomas and the expression is reduced or lost in sarcomatoid differentiation; together with the finding that (3) syndecan-1 correlates with WT1 immuno-expression, this suggests that syndecan-1 might relate to the differentiation state of mesothelial/mesothelioma cells; and (4) syndecan-1-positive tumours are associated with a longer survival (p = 0.02) than mesotheliomas with no or little syndecan-1 expression, on univariate analysis. These findings therefore indicate that syndecan-1 can be an important prognostic indicator in mesotheliomas and its loss may be important in the epithelial-mesenchymal transformation of mesothelioma cells.

Topics: Adenocarcinoma; Biomarkers, Tumor; Blotting, Western; Cell Differentiation; Chi-Square Distribution; DNA-Binding Proteins; Epithelium; Fibroblast Growth Factor 2; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Hyperplasia; Immunohistochemistry; Membrane Glycoproteins; Mesothelioma; Neovascularization, Pathologic; Precipitin Tests; Prognosis; Proteoglycans; Survival Rate; Syndecan-1; Syndecans; Transcription Factors; WT1 Proteins

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

Topics: Adenocarcinoma; Basement Membrane; Cell Differentiation; Chondroitin Sulfates; Eccrine Glands; Fluorescent Antibody Technique; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Proteoglycans; Sweat Gland Neoplasms

There is increasing evidence that the association of prostate carcinoma cells with the surrounding extracellular matrix is important for their growth. Proteoglycans (PGs) are components of matrix that form important associations with other molecules. Heparan sulfate proteoglycan (HSPG), for example, associates with several matrix components such as fibronectin, laminin, and basic fibroblast growth factor. The purpose of this study was to determine if human prostate carcinoma cells (PC-3) synthesized HSPG in tissue culture. Three areas of sulfate-labeled matrix were collected from PC-3 cells and analyzed: attachment sites, nonattachment cell surfaces, and media. Charged groups were separated by ion-exchange columns, and PGs identified by gel filtration chromatography after chemical and enzymatic treatment. The media fraction contained the greatest proportion of sulfated PGs (79.3%), of which 45.5% were HSPG. The greatest concentration of HSPG was in the attachment site fraction where 88.6% of PGs were HSPG, although only 11.7% of sulfated molecules were PGs. In nonattachment surfaces, only 19.6% were PGs, of which 19.4% were HSPG. When PGs were assessed for hydrophobic binding to octyl-Sepharose beads, only a proportion of HSPG in the media fraction contained hydrophobic domains (18.8%). In summary, PC-3 cells synthesize at least two types of HSPG, sorting them into different matrix compartments. The major PG in attachment sites is HSPG without a hydrophobic domain, while an HSPG in the media fraction has a hydrophobic domain. The localization of different types of HSPG may be functionally important and may be altered during the metastatic process for prostate carcinoma when in association with specific stroma.

Topics: Adenocarcinoma; Chromatography, Ion Exchange; Extracellular Matrix; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Male; Prostatic Neoplasms; Proteoglycans; Tumor Cells, Cultured

The status of the basement membrane in prostatic intraepithelial neoplasia (PIN) and adenocarcinoma is unsettled. Previous studies using antibodies directed against Type IV collagen have been hindered by intense staining around the stromal smooth muscle fibers, making interpretation of acinar staining difficult. We employed a monoclonal antibody to heparan sulfate proteoglycan (HSPG) to overcome this problem, recognizing that HSPG is present in the basement membrane of epithelial and endothelial cells, but not stromal smooth muscle cells. In 22 totally-embedded whole mount radical prostatectomies for adenocarcinoma which contained PIN, intense HSPG immunoreactivity was observed in the basement membrane of all normal and hyperplastic acini, 98% of acini with high grade PIN, and 100% of acini of well differentiated (Gleason score 5) adenocarcinoma; vessels served as the internal positive control, with consistent staining throughout each specimen. The extent of HSPG immunoreactivity in cancer decreased with increasing Gleason grade (measured as percent of acini staining, in 10% increments; p = 0.002). These findings indicate that HSPG is a consistent component of the basement membrane of benign, hyperplastic, and early neoplastic prostatic acini, and, unlike other extracellular matrix proteins such as type IV collagen, is not hindered by background staining around stromal smooth muscle cells. High grade PIN and well differentiated adenocarcinoma usually maintain an intact basement membrane, and loss of the basement membrane occurs with histologic dedifferentiation.

Topics: Adenocarcinoma; Aged; Antibodies, Monoclonal; Basement Membrane; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Male; Middle Aged; Neoplasm Staging; Neoplasms, Multiple Primary; Prostatic Intraepithelial Neoplasia; Prostatic Neoplasms; Proteoglycans

The prognosis for carcinoma of the pancreas is extremely poor. One of the characteristics of this tumor is its invasion of the surrounding tissues. Reduction of glycoprotein is considered to be conducive to invasion of the basement membrane by carcinoma cells. Heparan sulfate proteoglycan (HSPG), a kind of glycoprotein, is an important component of basement membrane. In this study, the relation between HSPG and carcinoma of the pancreas was examined by using the immunohistochemical method, and the survival rate of pancreatic adenocarcinoma was evaluated. We found that some carcinomas contained little or no HSPG. The poorer the differentiation of an adenocarcinoma of the pancreas, the lower was its content of HSPG. The level of HSPG was significantly different in carcinomatous and in noncarcinomatous cells. There was a close correlation among the content of HSPG, the degree of differentiation of carcinomas of the pancreas, and the survival time. HSPG seems to be useful in prognosis of adenocarcinoma of the pancreas.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Female; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Immunohistochemistry; Male; Middle Aged; Pancreatic Neoplasms; Prognosis; Proteoglycans; Survival Rate

Tissue samples from 30 patients with adenoid cystic carcinoma and 20 with adenocarcinoma of salivary gland origin were studied by immunohistochemical staining with specific antibodies to the four macromolecules that are present in normal basement membranes: type IV collagen, laminin, heparan sulfate proteoglycan, and entactin. In the adenoid cystic carcinoma samples, the four proteins were localized in different types of extracellular matrices in the tumor, namely pseudocystic spaces, hyaline stroma, and around tumor cell nests. The staining intensity was enhanced by pretreatment with hyaluronidase. The tumor cells of adenoid cystic carcinoma showed a tendency to proliferate with individual cells in contact with the basement membrane and to infiltrate through basement membrane-rich tissues, such as peripheral nerves, blood vessels, and skeletal muscles. In contrast, only circumferential staining of tumor cell nests was obtained in adenocarcinoma samples. The results suggest that adenoid cystic carcinoma is a tumor with affinity for basement membranes, and this basic feature is reflected in its histology and presumably in its biologic behavior. Immunostaining with antibodies to basement membrane proteins appears to be useful for differential diagnosis of some types of these two carcinomas.

Topics: Adenocarcinoma; Basement Membrane; Carcinoma, Adenoid Cystic; Collagen; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Immunohistochemistry; Laminin; Membrane Glycoproteins; Neoplasm Proteins; Proteoglycans

Double-label immunofluorescence was used to monitor basement-membrane composition and integrity in 22 human colon polyps, 36 adenocarcinomas and 2 metastases. Cryostat sections were stained with polyclonal anti-laminin anti-serum combined with monoclonal antibodies (MAbs) to all major basement-membrane components (laminin, entactin/nidogen, collagen type IV and large heparan sulfate proteoglycan), as well as to keratin 8. In all adenocarcinomas, including mucinous, basement membranes were altered more at the invasive front than in the parenchyma. The degree of this alteration was inversely correlated with the level of tumor differentiation. An uncoordinated loss of basement membrane components (dissociation of markers), previously described by us in rat colon adenocarcinomas, was also found in human tumors. In the great majority of adenocarcinomas a pronounced stromal reaction was seen. It was manifested by the presence of fibrillar deposits of basement-membrane components, mainly of collagen type IV and/or heparan sulfate proteoglycan. This reaction was never observed in polyps and may be derived from myofibroblasts reported to accumulate in colon cancer stroma. The combined use of antibodies to basement-membrane components and to a specific keratin may constitute an adequate immunohistochemical test for the presence of invasion, and may be useful in the histologic analysis of polyps, especially in dubious cases.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Basement Membrane; Blotting, Western; Collagen; Colon; Extracellular Matrix Proteins; Fluorescent Antibody Technique; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Intestinal Neoplasms; Intestinal Polyps; Keratins; Laminin; Lymphatic Metastasis; Membrane Glycoproteins; Proteoglycans

Two human colon carcinoma cell lines, HT29 and Caco-2 were co-cultured with fetal rat or human skin fibroblasts. Their morphological features, ultra-structural characteristics at the heterologous cell interface, and the deposition of basement-membrane molecules [laminin, type-IV collagen, heparan sulfate proteoglycan (HSPG)] at the epithelial-stromal junction were analyzed. The 2 cell lines behaved differently. HT29 cells did not spread on the fibroblasts and grew as clusters, while Caco-2 cells formed a monolayer over the fibroblastic feeder layer. Only the latter carcinoma cells exhibited cytoplasmic processes towards the fibroblasts and, after 5 days in co-cultures, a structured basement membrane (BM). The immunocytochemical analysis of the BM constituents revealed the absence of the molecules studied at the sites of heterologous contacts in the case of HT29 cells. In contrast, in the co-cultures comprising Caco-2 cells, laminin and type-IV collagen were progressively deposited in a polar fashion at the epithelial-fibroblastic interface which, however, remained devoid of HSPG molecules. Together with earlier data indicating a dual origin of the BM molecules located at the epithelial-fibroblastic interface in normal intestine, the present study shows that the cancer cells as well as the fibroblastic ones under the influence of carcinoma cells display an altered capacity to synthesize and/or secrete BM molecules. The extent of such abnormalities correlates with the differentiation of the cells. Finally, these modifications occur concomitantly with alterations in cell interactions which vary among cell lines.

Topics: Adenocarcinoma; Animals; Basement Membrane; Cell Adhesion; Cell Communication; Cell Differentiation; Cell Line; Cells, Cultured; Chondroitin Sulfate Proteoglycans; Collagen; Colonic Neoplasms; DNA Replication; Epithelium; Fetus; Fibroblasts; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Intercellular Junctions; Laminin; Microscopy, Electron; Rats; Skin; Skin Physiological Phenomena

Biosynthesis of glycosaminoglycans (GAGs) was studied in morphologically normal colonic mucosa, in peritumoral and tumoral areas, and in colorectal polyps of tumor-bearing patients. After GAG purification, overall biosynthesis was determined: the general trend was a decrease in GAG production in neoplastic colon, lowest GAG synthesis being observed in Dukes' stage C tumors. Separation by ion-exchange chromatography of various GAG species and further characterization revealed the presence of hyaluronic acid (HA) and heparan sulfate (HS) molecules in all specimens studied. Chondroitin-4 sulfate (CS4) was occasionally found in tumor samples. The relative proportion of HA and HS was modified in tumor tissue: i.e. increased HA and decreased HS were observed. Differences in DEAE-chromatographic behavior were obvious in pathological samples as compared to controls, the hydrodynamic form of HA and the charge density of HS being decreased. The latter could be attributed to undersulfatation of HS molecules. Immunocytochemical detection of HS proteoglycan molecules revealed regular and bright labelling at epithelial-stromal interface in control samples. In pathological samples, staining was patchy and discontinuous, showing large areas of basement membrane interruption.

Topics: Adenocarcinoma; Chondroitin Sulfate Proteoglycans; Colon; Colonic Polyps; Colorectal Neoplasms; Glycosaminoglycans; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Immunohistochemistry; Intestinal Mucosa; Rectum

The proteoglycans secreted by a malignant human breast cell line (MDA-MB-231) were compared with the corresponding proteoglycans from a normal human breast cell line (HBL-100). The physicochemical characteristics of these proteoglycans were established by hexosamine analysis, chemical and enzymatic degradations, and dissociative cesium chloride density gradient centrifugation, and by gel filtration before and after alkaline beta-elimination. Both cell lines secreted approximately 70% of the synthesized proteoglycans, which were composed of 20% heparan sulfate and 80% chondroitin sulfate proteoglycans. The MDA cell line secreted large hydrodynamic size (major) and small hydrodynamic size heparan sulfate proteoglycan. In contrast HBL cells secreted only one species having a hydrodynamic size intermediate to the above two. The chondroitin sulfate proteoglycans from MDA medium were slightly larger than the corresponding polymers from HBL medium. All proteoglycans except the small hydrodynamic size heparan sulfate proteoglycan from MDA medium were of high buoyant density. The proteoglycans of both cell lines contained significant proportions of disulfide-linked lower molecular weight components which were more pronounced in the proteoheparan sulfate polymers, particularly those from MDA medium, than in chondroitin sulfate proteoglycans. The glycosaminoglycans of heparan sulfate proteoglycans from MDA medium were more heterogeneous than those from HBL medium. The glycosaminoglycan chains of large hydrodynamic size heparan sulfate proteoglycans from MDA medium were larger in size than those from HBL medium while small hydrodynamic size heparan sulfate proteoglycans contained shorter glycosaminoglycan chains. In contrast to the glycosaminoglycans derived from chondroitin sulfate proteoglycans of both MDA and HBL medium were comparable in size. The heparan sulfate as well as chondroitin sulfate proteoglycans of both cell lines contained both neutral (di- and tetrasaccharides) and sialylated (tri- to hexasaccharides) O-linked oligosaccharides.

Topics: Adenocarcinoma; Breast; Breast Neoplasms; Carbohydrate Conformation; Cell Line; Centrifugation, Density Gradient; Chondroitin Sulfates; Chromatography, Gel; Disulfides; Epithelium; Female; Galactosamine; Glucosamine; Glycosaminoglycans; Heparitin Sulfate; Humans; Molecular Weight; Oligosaccharides; Proteoglycans

TA3 murine ascites adenocarcinoma cells were compared for their ability to release radioactive glucosamine and 35SO4-labeled glycoproteins and glycosaminoglycans into the culture medium. Both TA3-Ha and TA3-St cells contained cell-surface heparan sulfate that was released into culture, but not chondroitin sulfate. Both cells released a membranous aggregate of labeled components from the cell surface and hyaluronic acid from inside the cells that fractionated in the void volume of Sepharose CL-4B. This void-volume fraction from the TA3-Ha cells contained glucosamine-labeled epiglycanin at a higher concentration relative to other glucosamine-labeled components than that found on plasma membranes. Glycoproteins associated with epiglycanin found on the cell surface, as well as released into culture medium, contained sulfate that could not be removed by chondroitinase ABC, heparinase, or keratinase. Kinetic analysis of the glucosamine-labeled material released from TA3-Ha cells indicated that hyaluronic acid was released rapidly with a 45-min half-life, whereas the other membranous components were released much more slowly.

Topics: Adenocarcinoma; Animals; Carbon Radioisotopes; Cell Line; Cell Membrane; Glucosamine; Glycoproteins; Glycosaminoglycans; Hemagglutination Inhibition Tests; Heparitin Sulfate; Leucine; Mice; Sulfur Radioisotopes; Tritium

The sulfated glycosaminoglycan (SGAG) composition of gastric mucosa from 24 patients with chronic superficial gastritis, 2 patients with adenocarcinoma and 36 normal subjects is reported. The mucosa was obtained by endoscopic biopsy and after histopathological examination the SGAG were extracted and characterized. Three different SGAG were isolated: chondroitin 4,6-sulfate, dermatan sulfate and heparan sulfate. Their relative concentrations for the different groups were submitted to analysis of variance by Scheffe's method. Different SGAG compositions were observed in two gastric regions (antrum and body), in chronic superficial gastritis, in adenocarcinoma and in two age groups (less than 40 years and greater than 40 years). These and other results suggest that these macromolecules might be involved in the processes of cell division and aging.

Topics: Adenocarcinoma; Adult; Aging; Chondroitin Sulfates; Dermatan Sulfate; Gastric Mucosa; Gastritis; Glycosaminoglycans; Heparitin Sulfate; Humans; Stomach Neoplasms

This unit has in the past evaluated the Nb rat prostatic adenocarcinoma model with respect to chemotherapies. Recently, this unit has been evaluating agents that may have a role in decreasing metastatic rate. The three androgen-insensitive tumors, Nb Pr A.I. I, II, III, have been evaluated herein. Agents that have been used include indomethacin, heparin, heparin plus cortisone, and heparan sulfate (SP54). It has been shown that these agents do play a role in reducing the metastasis. In evaluation of Nb Pr A.I, I, control animals had a metastatic rate of 57%. In treatment with indocin, only 21% of the animals, three of 14 animals treated, had a metastasis, and treatment with heparin and cortisone resulted in one of 14 animals having metastasis. Similar observations were seen when treatment with SP54 and heparin was evaluated in the Nb Pr A.I. III; 18 and 27% of treatment group animals had metastasis, whereas 55% of control groups had metastasis. Similarly, in the Nb Pr A.I. II evaluation, control animals having a metastatic rate of 43% had heparin plus cortisone and heparin alone, and this particular tumor model revealed complete resolution with no animals having metastatic disease. The majority of these agents have not effected tumor volume in terms of reduction as much as the best chemotherapeutic agents in this model system include cyclophosphamide and cis-platinum.

Topics: Adenocarcinoma; Animals; Antineoplastic Agents; Cortisone; Disease Models, Animal; Drug Therapy, Combination; Heparin; Heparitin Sulfate; Indomethacin; Male; Neoplasm Metastasis; Prostatic Neoplasms; Rats; Rats, Inbred Strains

Using a modified papain digestion cetylpyridinium salt precipitation method, glycosaminoglycans were isolated from 21 mesotheliomas, 34 primary lung carcinomas, 12 carcinomas of other sites, and 7 soft tissue sarcomas. Qualitatively, hyaluronic acid (HA) was present in 20 of 21 mesotheliomas, about half of the primary lung adenocarcinomas, and all of the soft tissue sarcomas. On the average, HA constituted 45% of the total glycosaminoglycans in the mesotheliomas and 28% of the total in the lung cancers. Quantitatively, mesotheliomas contained statistically greater amounts (mean value, 0.74 mg/g) of HA than primary lung adenocarcinomas (mean value, 0.08 mg/g), but were not statistically different from soft tissue sarcomas (mean value, 2.01 mg/g) or primary ovarian serous neoplasms (mean value, 0.92 mg/g). The study concludes that, contrary to previous reports, HA is neither the sole nor the predominant glycosaminoglycan in most mesotheliomas, but, given the proper clinical and histologic setting, the finding of sufficiently high levels (greater than 0.4 mg/g dry tissue extract) supports the diagnosis of mesothelioma when the alternative diagnosis is primary adenocarcinoma of lung.

Topics: Adenocarcinoma; Carcinoma; Chondroitin Sulfates; Dermatan Sulfate; Diagnosis, Differential; Electrophoresis, Cellulose Acetate; Female; Glycosaminoglycans; Heparitin Sulfate; Humans; Hyaluronic Acid; Lung Neoplasms; Mesothelioma; Ovarian Neoplasms; Sarcoma

The influence of fixed fibroblasts on the glycosaminoglycan (GAG) synthesis of gastric carcinoma cells was examined by incubation along with [3H]glucosamine. In well-differentiated adenocarcinoma cells, the amount of 3H-GAG in the interface material between the carcinoma cells and the fixed fibroblasts was much larger (about twenty times) than in the interface between the carcinoma cells and the bare culture plates, and 3H-GAG consisted mainly of heparan sulfate, with a small amount of dermatan sulfate and chondroitin sulfate. On the other hand, in poorly differentiated carcinoma cells, the amount of 3H-GAG in the interface material produced by the carcinoma cells on the fibroblast was almost the same as on the bare culture dish. In a conventional monolayer culture, well-differentiated adenocarcinoma cells produced a much greater amount of GAG, consisting mainly of dermatan sulfate, chondroitin sulfate and heparan sulfate, than poorly differentiated carcinoma cells. Almost the same amount of hyaluronic acid was secreted into the medium by both types of carcinoma cells.

Topics: Adenocarcinoma; Cell Adhesion; Cell Communication; Cell Differentiation; Cell Division; Cell Line; Chondroitin Sulfates; Dermatan Sulfate; Fibroblasts; Glycosaminoglycans; Heparitin Sulfate; Humans; Stomach Neoplasms

A transplantable colorectal adenocarcinoma and the normal colonic mucosa derived from rats of ACI/N strain were digested with pronase, and the glycosaminoglycan fractions were obtained by fractionation with cetylpyridinium chloride. The glycosaminoglycan fraction derived from the adenocarcinoma contained substantial amounts of chondroitin sulfate A/C, dermatan sulfate, heparan sulfate, and hyaluronic acid, whereas chondroitin sulfate A/C and dermatan sulfate were undetectable in that derived from the normal colonic mucosa. An increment in the heparan sulfate content was also apparent in the adenocarcinoma, while the level of hyaluronic acid appeared to be unchanged. Analyses of the extract of the tumor tissue with 5mM ethylenediaminetetraacetate (pH 7.0) indicated that heparan sulfate was present largely, if not completely, in the form of proteoglycan.

Topics: Adenocarcinoma; Animals; Chemical Fractionation; Chondroitin Sulfates; Colon; Colonic Neoplasms; Dermatan Sulfate; Glycosaminoglycans; Heparitin Sulfate; Hyaluronic Acid; Mucous Membrane; Neoplasms, Experimental; Rats; Rats, Inbred ACI; Rectal 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 from normal colonic mucosa and colons with a variety of inflammatory diseases, as well as benign and malignant neoplasms were analyzed. Normal colonic mucosa contains predominantly chondroitin sulfates and dermatan sulfate. Increases in the levels of hyaluronic acid and heparan sulfate, as well as substantial increases in the amount of total glycosaminoglycans were characteristic of invasive colonic adenocarcinoma. Lesser elevations in the amount of total glycosaminoglycans and hyaluronic acid and heparan sulfate were present in neonatal colonic mucosa, villous adenoma, ulcerative colitis, and mucosa adjacent to carcinoma. The degree of elevation was proportional to the dysplastic potential. Since dysplastic lesions have scant connective tissue, the epithelial component of colonic neoplasms may contribute to these neoplasm-related alterations in glycosaminoglycan composition.

Topics: Adenocarcinoma; Adult; Aged; Chondroitin Sulfates; Colitis; Colon; Colonic Neoplasms; Dermatan Sulfate; Female; Glycosaminoglycans; Heparitin Sulfate; Humans; Hyaluronic Acid; Intestinal Mucosa; Male; Middle Aged

Normal, transitional, and carcinoma areas of five colons resected for carcinoma were examined morphologically, histochemically, and biochemically. The transitional area contained a larger amount of non-sulphated acid mucin than the normal mucosa as verified histochemically. Normal mucosa contained mainly sulphated mucin. The hexosamine-containing macromolecules present in different areas were isolated and characterized. They were divided into the following groups: 1) acid glycosaminoglycans, 2) high-molecular-weight glycopeptides, and 3) low-molecular weight glycopeptides. The concentration of the total hexosamine-containing material was in the carcinoma area twice as high as in normal areas. Acid glycosaminoglycans were identified as hyaluronate, heparan sulphate, dermatan sulphate, and chondroitin 4-(6)-sulphate. Their concentraitons were found to increase from normal to transitional and from transitional to carcinoma areas. The high-molecular-weight glycopeptide was composed of fucose, galactose, glucosamine, galactosamine, sialic acid, and variable amounts of sulphate. The sulphation degree of the glycopeptide was higher in normal mucosa than in transitional or carcinoma areas: The low-molecular-weight glycopeptides consisted of about a half of the total hexosamine-containing substances. The concentration of saline-insoluble fraction of the low-molecular-weight glycopeptides was in transitional areas about two times, and in carcinoma areas about four times, higher than in normal mucosa.

Topics: Adenocarcinoma; Chondroitin Sulfates; Colonic Neoplasms; Dermatan Sulfate; Fucose; Galactosamine; Galactose; Glucosamine; Glycopeptides; Glycosaminoglycans; Heparitin Sulfate; Hexosamines; Histocytochemistry; Humans; Hyaluronic Acid; Intestinal Mucosa; Macromolecular Substances; Molecular Weight; Mucins; Sialic Acids; Sulfates

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

The polysaccharide composition of the human gallbladder well was studied in carcinomas and metaplastic changes of various degrees, and the results obtained were compared with those for the normal material previously presented (Terho, T., and Laitio, M. Biochim. Biophys. Acta 338: 135, 1974). Elevated amounts of acid connective tissue polysaccharides (heparitin and dermatan sulfates as well as chondroitin 4- or 6-sulfate, or both, could be observed in carinomas. In histochemical stainings it was found that in carcinomas and in the two specimens classified as group III (containing the most extensive metaplastic changes at disposal), the intracellular mucin was mainly neutral or nonsulfated acidic. The amounts of sulfated mucin were relatively insignificant. This mucin polysaccharide material was isolated and its composition was determined. It was observed to be large polysaccharide material was isolated and its composition was determined. It was observed to be large molecular (approximate molecular sizes 1 to 2 times 10-6), and to be composed of fucose, galactose, glucosamine, and galactosamine as well as small amounts of sialic acid. The basic structure of these polysaccharides is thus similar to that of normal sulfated mucin. The almost total absence of acid groups, however, causes the polysaccharide material in question to stain in a manner identical with neutral mucin when investigated with histochemical methods. The carcinomas also contained some sulfomucin; its proportion, however, was small as compared with the amounts of nonsulfated acid and neutral mucin in biochemical characterization. A small molecular polysaccharide fraction, assumed to originate in membrane-bound glycoproteins, was isolated from the insoluble gallbladder tissue residue. The proportion of this fraction was larger in carcinomas than in normal material. This rise as well as the rise in the quantity of acid connective tissue polysaccharides is presumably due to the large number of cells in the carcinoma tissue as well as to fibrosis.

Topics: Adenocarcinoma; Cetylpyridinium; Chemical Precipitation; Chondroitin; Dermatan Sulfate; Fucose; Galactosamine; Galactose; Gallbladder Neoplasms; Glucosamine; Heparitin Sulfate; Humans; Metaplasia; Mucins; Mucous Membrane; Polysaccharides; Sialic Acids; Solubility; Staining and Labeling