Inhibition of heparanase activity and tumor metastasis by laminarin sulfate and synthetic phosphorothioate oligodeoxynucleotides

Miao, Hua‐Quan; Elkin, Michael; Aingorn, Elena; Ishai-Michaeli, Rivka; Stein, C. A.; Vlodavsky, Israël

doi:10.1002/(sici)1097-0215(19991029)83:3<424::aid-ijc20>3.0.co;2-l

Cited by 162 publications

(96 citation statements)

References 19 publications

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“…Similar results were obtained with sulfate-labeled nuclear preparation of C6 rat glioma cells (not shown). Laminaran sulfate (5 mg/ml), a potent inhibitor of heparanase activity and experimental metastasis, 42 inhibited the degradation of nuclear HS by heparanase (Figure 4), as previously demonstrated for HS in the ECM. 42 …”

Section: Nuclear Heparan Sulfate Is a Substrate For Nuclear Heparanasesupporting

confidence: 78%

Human heparanase nuclear localization and enzymatic activity

Schubert¹,

Ilan²,

Shushy³

et al. 2004

Laboratory Investigation

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

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Section: Nuclear Heparan Sulfate Is a Substrate For Nuclear Heparanasesupporting

confidence: 78%

Human heparanase nuclear localization and enzymatic activity

Schubert¹,

Ilan²,

Shushy³

et al. 2004

Laboratory Investigation

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“…31 It therefore appears that vascular-, and immune cellsderived heparanase, contributes to the early phases of an inflamed condition, 31 whereas prolonged exposure to pro-inflammatory cytokines results in heparanase induction by the colonic epithelium, contributing to the sustained, chronic nature of the disease. Interestingly, un-fractionated as well as low molecular weight heparin (enoxaparin), which exhibit a strong inhibitory activity towards heparanase, [32][33][34][35] have proven efficacious in ulcerative colitis and Crohn's disease patients, [36][37][38] suggesting that heparanase is actively involved in these pathologies and thus may be considered as a target for the development of anti-inflammatory therapies. …”

Section: Discussionmentioning

confidence: 99%

Heparanase upregulation by colonic epithelium in inflammatory bowel disease

et al. 2007

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Heparanase is an endo-b-D-glucuronidase capable of cleaving heparan sulfate (HS) side chains at a limited number of sites, yielding HS fragments of still appreciable size (B5-7 kDa). Heparanase activity has long been detected in a number of cell types and tissues. Importantly, heparanase activity correlated with the metastatic potential of tumor-derived cells, attributed to enhanced cell dissemination as a consequence of HS cleavage and remodeling of the extracellular matrix barrier. Similarly, heparanase activity was implicated in neovascularization, inflammation and autoimmunity, involving migration of vascular endothelial cells and activated cells of the immune system. The involvement of heparanase in inflammatory processes of the gastrointestinal tract has not been examined. Here, we utilized immunohistochemical analysis to investigate heparanase expression in acute and chronic inflammatory conditions. Heparanase expression was not detected in specimens derived from normal colon tissue. In contrast, strong heparanase staining was observed in Crohn's disease and ulcerative colitis, but not in infectious colitis. Interestingly, heparanase staining was primarily observed in epithelial rather than immune cells. Importantly, un-fractionated as well as low molecular weight heparin (enoxaparin), which exhibit a strong inhibitory activity towards heparanase, have proven efficacious in ulcerative colitis and Crohn's disease patients, suggesting that heparanase is actively involved in these pathologies and thus may be considered as a target for the development of anti-inflammatory therapies. The mammalian endoglycosidase heparanase is the predominant enzyme that degrades heparan sulfate (HS) side chains of heparan sulfate proteoglycans (HSPG), the dominant proteoglycan in the extracellular matrix (ECM) and cell surfaces. 1-3 HSPG consist of a protein core to which HS side chains are covalently attached. These complex macromolecules are highly abundant in the ECM and are thought to play an important structural role, contributing to ECM integrity and insolubility. 4,5 Traditionally, heparanase activity was well correlated with the metastatic potential of tumor-derived cells, attributed to enhanced cell dissemination as a consequence of HS cleavage and remodeling of the ECM barrier. 6-9 A proof-of-concept of this notion has recently been established by using a specific antiheparanase ribosyme and siRNA methodology, 10 clearly implicating heparanase activity as a critical requisite for metastatic spread. Similarly, heparanase was implicated in cell dissemination associated with inflammation and autoimmunity. [11][12][13] Furthermore, heparanase activity can liberate a variety of HSPG-bound biological mediators, including cytokines and chemokines such as interferon (IFN)-g, MIP-1b, RANTES and interleukins, thus contributing to the regulation of inflammation and other immune responses. 14 Heparanase expression by the gastrointestinal tract has been documented by employing immunostaining, RT-PCR and heparanase activity analy...

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“…Some reports have demonstrated that heparanase activity can be blocked with various nonanticoagulant heparin derivatives and related sulfated negatively charged molecules. [18][19][20][21] These factors that decrease heparanase expression and/or inhibit heparanase enzymatic activity in cancer cells significantly reduce their metastatic properties, signifying the importance of heparanase in cancer cell spread. Interestingly, no significant similarity or homology was observed between the human and murine heparanase sequences and any other reported protein.…”

Section: Discussionmentioning

confidence: 99%

“…A Volume of 7 ml of total RNA isolated from approximately 10 000 cells were mixed with 1 ml of oligo-(dT) [12][13][14][15][16][17][18] primer, 1 ml of random hexamers primer (N 6 ), and 1 ml of 10 mM dNTPs in a total volume of 10 ml. They were heat denaturated at 651C for 5 min, and then chilled in ice.…”

Section: Rt-pcr Of Microdissected Cellsmentioning

confidence: 99%

Positive association of heparanase expression with tumor invasion and lymphatic metastasis in gastric carcinoma

Wang

Jiang

et al. 2005

Modern Pathology

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Tumor invasion and metastasis are the most common causes of death in gastric carcinoma. Human heparanase influences tumor invasiveness and angiogenesis. Analysis of its expression in gastric carcinoma has been hindered by our inability to procure pure cancer cells from heterogeneous tissue. In the present study, we analyzed heparanase expression in human primary and metastatic gastric carcinoma cells as well as in paired normal gastric epithelial cells by laser capture microdissection coupled with reverse transcriptionpolymerase chain reaction (RT-PCR). Tumor tissues, metastatic lymph nodes, and apparently uninvolved normal gastric tissues were collected from 30 patients who had undergone gastrectomy with radical lymph node dissection for gastric carcinoma without preoperative treatment. Bulk tissues and laser capture microdissected cell groups were separately subjected to RT-PCR analysis with heparanase-specific primers. For bulk tissues, heparanase-specific transcripts were detectable in all primary tumor tissues, metastatic lymph nodes, and almost all matching normal tissues. RT-PCR analysis after laser capture microdissection showed no detectable heparanase expression in matching normal epithelial cell groups. Of the laser capture microdissected primary gastric carcinoma cells, 47% (14/30) were heparanase positive. Expression was closely associated with greater tumor invasiveness, including Borrmann gross type and depth of wall infiltration. For metastatic cell groups dissected from lymph nodes, 95% showed clear heparanase expression. Furthermore, the extent of lymphatic spread was directly correlated to heparanase expression at the primary site. In conclusion, laser capture microdissection coupled with RT-PCR is a reliable approach for molecular analysis of heparanase expression in gastric carcinoma. Heparanase may facilitate invasion and metastasis of gastric carcinoma cells. Keywords: gastric carcinoma; laser capture microdissection; lymphatic metastasis; heparanase expression It is generally accepted that the invasion of the basement membrane and extracellular matrix is one of the critical steps for cancer cell metastasis. 1 Heparan sulfate proteoglycans, composed of a protein core covalently linked to heparan sulfate side-chains, represent a principal component of the extracellular matrix and, in particular, basement membranes. Cell surface heparan sulfate can also serve as coreceptors, along with the other cell surface molecules, to form functional receptor complexes that facilitate signal transduction. 2 Furthermore, heparan sulfate chains bind a large number of bioactive molecules and regulate their availability and function. These include growth factors, chemokines, cytokines, and enzymes that are essential for angiogenesis, cell adhesion, and locomotion. 3 Accordingly, enzymatic degradation of heparan sulfate may play a critical role in cancer cell invasion and metastasis. Heparanase is an endoglycosidase that cleaves heparan sulfate side chains of heparan sulfate proteoglycans at a limited number ...

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Inhibition of heparanase activity and tumor metastasis by laminarin sulfate and synthetic phosphorothioate oligodeoxynucleotides

Cited by 162 publications

References 19 publications

Human heparanase nuclear localization and enzymatic activity

Human heparanase nuclear localization and enzymatic activity

Heparanase upregulation by colonic epithelium in inflammatory bowel disease

Positive association of heparanase expression with tumor invasion and lymphatic metastasis in gastric carcinoma

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