heparitin-sulfate and Kidney-Neoplasms

Previous studies have demonstrated that the glycosaminoglycans (GAGs) heparan sulfate (HS) and hyaluronic acid (HA) are mechanosensors for interstitial flow on cancer cells. The proteins that link the GAGs to the cancer cell for mechanotransduction, however, are not known.. To assess whether the HS proteoglycan core proteins, Glypican-1 and Syndecan-1, or the HA receptor, CD44, provides the mechanical linkage to the cell.. The highly metastatic renal carcinoma cell line (SN12L1) and its companion low metastatic cell line (SN12C) were analyzed by Western blot, siRNA, and a 3-dimensional interstitial flow migration assay.. There was significant elevation of Glypican-1 protein expression in the SN12L1 cells relative to the SN12C cells while there were no significant differences in Syndecan-1 or CD44. Knock down of Glypican-1 by siRNA completely blocked flow induced migration in SN12L1 cells. MAPK inhibitors also blocked flow induced migration in SN12L1 cells.. Glypican-1 provides the mechanical linkage from HS (the flow sensor) to the SN12L1 cell where mechanotransduction leading to the enhancement of migration (metastasis) occurs. MAPKs downstream of Glypican-1 propagate the signal. The HS, Glypican-1, MAPK signaling axis suggests opportunities for pharmaceutical intervention.

Topics: Carcinoma, Renal Cell; Cell Line, Tumor; Cell Movement; Extracellular Fluid; Extracellular Signal-Regulated MAP Kinases; Glycocalyx; Glypicans; Heparitin Sulfate; Humans; Hyaluronan Receptors; Kidney Neoplasms; Mechanotransduction, Cellular; Neoplasm Metastasis; Syndecan-1

Glycosaminoglycans (GAGs) are heterogeneous, linear, highly charged, anionic polysaccharides consisting of repeating disaccharides units. GAGs have some biological significance in cancer progression (invasion and metastasis) and cell signaling. In different cancer types, GAGs undergo specific structural changes. In the present study, in depth investigation of changes in sulfation pattern and composition of GAGs, heparan sulfate (HS)/heparin (HP), chondroitin sulfate (CS)/dermatan sulfate and hyaluronan (HA) in normal renal tissue (NRT) and renal cell carcinoma tissue (RCCT) were evaluated. The statistical evaluation showed that alteration of the HS (HSNRT = 415.1 ± 115.3; HSRCCT = 277.5 ± 134.3), and CS (CSNRT = 35.3 ± 12.3; CSRCCT = 166.7 ± 108.8) amounts (in ng/mg dry tissue) were statistically significant (p < 0.05). Sulfation pattern in NRT and RCCT was evaluated to reveal disaccharide profiles. Statistical analyses showed that RCCT samples contain significantly increased amounts (in units of ng/mg dry tissue) of 4SCS (NRT = 25.7 ± 9.4; RCCT = 117.1 ± 73.9), SECS (NRT = 0.7 ± 0.3; RCCT = 4.7 ± 4.5), 6SCS (NRT = 6.1 ± 2.7; RCCT = 39.4 ± 34.7) and significantly decreased amounts (in units of ng/mg dry tissue) of NS6SHS (RCCT = 28.6 ± 6.5, RCCT = 10.2 ± 8.0), NS2SHS (RCCT = 44.2 ± 13.8; RCCT = 27.2 ± 15.0), NSHS (NRT = 68.4 ± 15.8; RCCT = 50.4 ± 21.2), 2S6SHS (NRT = 1.0 ± 0.4; RCCT = 0.4 ± 0.3), and 6SHS (NRT = 60.6 ± 17.5; RCCT = 24.9 ± 12.3). If these changes in GAGs are proven to be specific and sensitive, they may serve as potential biomarkers in RCC. Our findings are likely to help us to show the direction for further investigations to be able to bring different diagnostic and prognostic approaches in renal tumors.

Topics: Biomarkers, Tumor; Carcinoma, Renal Cell; Chondroitin Sulfates; Dermatan Sulfate; Heparitin Sulfate; Humans; Kidney Neoplasms

Fibroblast growth factors (FGFs) play multiple roles during development and in adult tissues as paracrine regulators of growth and differentiation. FGFs signal through transmembrane receptor tyrosine kinases, but heparan sulfate is also required for signaling by members of the FGF family. In addition, heparan sulfate may be involved in determining tissue distribution of FGFs. Using biotinylated FGF-2 and FGF-7 (KGF) as probes, we have identified specific interactions between FGFs and heparan sulfates in human tissues. Both FGF species bind to tissue mast cells and to epithelial cell membranes. Binding to basement membrane heparan sulfate is tissue source dependent and specific. Although FGF-2 strongly binds to basement membrane heparan sulfate in skin and most other tissue sites examined, FGF-7 fails to bind to basement membrane heparan sulfate in most locations. However, in subendothelial matrix in blood vessels and in the basement membrane of a papillary renal cell carcinoma, strong FGF-7 binding is seen. In summary, distinct and specific affinities of heparan sulfates for different FGFs were identified that may affect growth factor activation and local distribution. Heparan sulfate may have a gatekeeper function to either restrict or permit diffusion of heparin-binding growth factors across the basement membrane.

Topics: 3T3 Cells; Animals; Basement Membrane; Carcinoma, Renal Cell; Epidermis; Fibroblast Growth Factor 10; Fibroblast Growth Factor 2; Fibroblast Growth Factor 7; Fibroblast Growth Factors; Growth Substances; Heparitin Sulfate; Humans; Kidney Neoplasms; Kidney Tubules; Mice; Protein Binding; Skin

In the present study we provide evidence for the involvement of N-CAM in the spreading of human renal cell carcinomas (RCC) through the interaction with the subendothelial matrix. We found that in tumor cell lines derived from human RCC the increase of growth rate and the loss of adhesiveness to inert substrate were accompanied by N-CAM expression and by the appearance of specific binding to endothelial heparan sulfate. Indeed, the adhesion of tumor cells to human endothelial cells and heparan sulfate in vitro was inhibited by monoclonal antibodies able to bind and inactivate N-CAM and was abrogated by endothelial cell treatment with heparitinase. Furthermore, when the renal epithelial cell line COS7 was transfected with a cDNA coding for N-CAM a significant increase in the ability to bind both endothelium and heparan sulfate in vitro was observed. Of note, HS complexed with epithelial growth factor could enhance the proliferation of RCC-derived tumor cells; this effect was also achieved by cross-linking of N-CAM at the surface of tumor cells, suggesting that N-CAM could transduce an activation signal across the cell membrane. This was also supported by the finding that N-CAM cross-linking induced a strong calcium mobilization from internal stores and opening of surface calcium channels in such tumor cells. N-CAM was detectable in vivo at the tumor site in the areas of active proliferation, as judged by the coexpression of Ki67 nuclear antigen, and heparan sulfate was present in the wall of blood vessels in the proximity of the tumor. These findings would suggest that growing kidney tumors might use N-CAM to bind the subendothelial matrix and complexed growth factors during tissue invasion and spreading.

Topics: Carcinoma, Renal Cell; Cell Adhesion; Cell Adhesion Molecules, Neuronal; Cell Division; Endothelium, Vascular; Extracellular Matrix Proteins; Heparitin Sulfate; Humans; Integrin alpha1; Integrin beta1; Integrins; Ki-67 Antigen; Kidney Neoplasms; Neoplasm Metastasis; Neoplasm Proteins; Nuclear Proteins; Signal Transduction

The glycosaminoglycans of the tumor mass and from the urine of patients with a nephroblastoma of embryonic origin (Wilms' tumor) and hypernephroma were analyzed. The urine of patients with Wilms' tumors prior to treatment, and two patients with metastasis contained high levels of hyaluronic acid (2-5 mg/l of urine) when compared to patients after surgery or chemotherapy where the content of hyaluronic acid was less than 0.1 mg/l. Urine of patients with hypernephroma and normal individuals contained even smaller amounts of hyaluronic acid. Normal kidneys contain mainly dermatan sulfate and heparan sulfate, while the hypernephroma and Wilms' tumor contain substantial amounts of chondroitin sulfate. The amount of glycosaminoglycans isolated from Wilms' tumor and hypernephroma were 10 times and 3 times, respectively, greater than normal kidneys. The amounts of hyaluronic acid in Wilms' tumor varied from 56 to 73% whereas normal kidneys contained about 13%. Chondroitin sulfate was also increased in Wilms' tumor and hypernephroma. It corresponded to 11% and 42%, respectively, of the total glycosaminoglycans. These and other findings indicate that the glycosaminoglycans of Wilms' tumors resemble those present during embryonic development of normal tissues whereas those in hypernephroma are typical of other carcinomas of different origins.

Topics: Carcinoma, Renal Cell; Dermatan Sulfate; Disaccharides; Glycosaminoglycans; Heparitin Sulfate; Humans; Hyaluronic Acid; Kidney Neoplasms; Wilms Tumor

In an experimental series of preneoplastic lesions and neoplasms induced in the rat kidney by dimethylnitrosamine, four of the 54 tumors showed particular morphological features that allowed them to be grouped separately. The leading characteristic of these lesions was an abundant PAS-positive matrix in which the tumor cells were embedded. The demonstration of variable amounts of laminin, heparan sulfate proteoglycan, collagen types IV and V in the tumor matrix provided circumstantial evidence that it consisted of basement membrane material. Since the tumor cells were in intimate contact with the matrix and no fibroblasts were present, it was assumed that the matrix was a product of the tumor cells. Until now, no similar renal tumors have been described in laboratory animals and no human equivalent is known.

Topics: Animals; Basement Membrane; Collagen; Dimethylnitrosamine; Epithelium; Female; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Immunohistochemistry; Kidney Neoplasms; Laminin; Periodic Acid-Schiff Reaction; Proteoglycans; Rats; Rats, Wistar; Staining and Labeling