heparitin-sulfate and Stomach-Neoplasms

Gastric cancer is a dominating cause of cancer-associated mortality with limited therapeutic options. Here, we show that syndecan-4 (SDC4), a transmembrane proteoglycan, is highly expressed in intestinal subtype gastric tumors and that this signature associates with patient poor survival. Further, we mechanistically demonstrate that SDC4 is a master regulator of gastric cancer cell motility and invasion. We also find that SDC4 decorated with heparan sulfate is efficiently sorted in extracellular vesicles (EVs). Interestingly, SDC4 in EVs regulates gastric cancer cell-derived EV organ distribution, uptake, and functional effects in recipient cells. Specifically, we show that

Topics: Heparitin Sulfate; Humans; Neoplasm Invasiveness; Stomach Neoplasms; Syndecan-4

Heparan sulfate (HS) proteoglycans (HSPGs) are abundant glycoconjugates in cells' glycocalyx and extracellular matrix. By acting as scaffolds for protein-protein interactions, HSPGs modulate extracellular ligand gradients, cell signaling networks, and cell-extracellular matrix crosstalk. Aberrant expression of HSPGs and enzymes involved in HSPG biosynthesis and processing has been reported in tumors, with impact in cancer cell behavior and tumor microenvironment properties. However, the roles of specific glycosyltransferases in the deregulated biosynthesis of HSPGs are not fully understood. In this study, we established glycoengineered gastric cancer cell models lacking either exostosin-like glycosyltransferase 2 (EXTL2) or EXTL3 and revealed their regulatory roles in both HS and chondroitin sulfate (CS) biosynthesis and structural features. We showed that EXTL3 is key for initiating the synthesis of HS chains in detriment of CS biosynthesis, intervening in the fine-tuned balance of the HS/CS ratio in cells, while EXTL2 functions as a negative regulator of HS biosynthesis, with impact over the glycoproteome of gastric cancer cells. We demonstrated that KO of EXTL2 enhanced HS levels along with concomitant upregulation of Syndecan-4, which is a major cell surface carrier of HS. This aberrant HS expression profile promoted a more aggressive phenotype, characterized by higher cellular motility and invasion, and impaired activation of Ephrin type-A 4 cell surface receptor tyrosine kinase. Our findings uncover the biosynthetic roles of EXTL2 and EXTL3 in the regulation of cancer cell GAGosylation and proteoglycans expression and unravel the functional consequences of aberrant HS/CS balance in cellular malignant features.

Topics: Cell Movement; Glycosyltransferases; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Membrane Proteins; N-Acetylglucosaminyltransferases; Stomach Neoplasms; Tumor Microenvironment

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

Human C21orf63 is a type-1 transmembrane protein of hitherto unknown function, with two repeats of putative 'galactose-binding lectin domains'. By using glycan microarray analysis and other assays, we found that human C21orf63 interacts with heparin and to a lesser extent with heparan sulphate. The C-terminal galactose-binding lectin domain of C21orf63 is necessary for heparin binding. The inability of other human proteins with galactose-binding lectin domains to interact with heparin suggests that heparin binding is a unique property of C21orf63. Results of real-time polymerase chain reaction and tissue immunostaining imply that C21orf63 is expressed on epithelia of various human tissues.

Topics: Cloning, Molecular; Databases, Protein; Epithelial Cells; Fluorescent Antibody Technique; Galectins; Gastric Mucosa; Gene Expression; Glycosaminoglycans; Heparin; Heparitin Sulfate; Humans; Immobilized Proteins; Lectins; Membrane Proteins; Microarray Analysis; Microscopy, Fluorescence; Mutant Proteins; Organ Specificity; Protein Binding; Protein Structure, Tertiary; Recombinant Fusion Proteins; Reverse Transcriptase Polymerase Chain Reaction; Stomach Neoplasms

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

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

The glycosaminoglycan (GAG) content of stomach carcinoma tissue was compared with that of non-neoplastic mucosa. GAG synthesis was also studied, by an analysis of 35S-labelled material after incubation of tissue segments in medium containing 35SO4. No significant difference was found between the amount of GAG and its components in the medullary carcinoma tissue and in non-neoplastic mucosa, but GAG synthesis of the carcinoma tissue was at a much higher rate than that of the non-neoplastic mucosa. In the autoradiograph, high 35S uptake in the carcinoma cells was observed. The GAG content of the scirrhous-carcinoma tissue was about twice that of medullary carcinoma.

Topics: Chondroitin Sulfates; Dermatan Sulfate; Gastric Mucosa; Glycosaminoglycans; Heparitin Sulfate; Humans; Hyaluronic Acid; Stomach Neoplasms