heparitin-sulfate and Pleural-Effusion--Malignant

While malignant pleural effusion (MPE) is a common and significant cause of morbidity in patients with cancer, current treatment options are limited. Human heparanase, involved in angiogenesis and metastasis, cleaves heparan sulfate (HS) side chains on the cell surface.. To explore the coagulation milieu in MPE and infectious pleural effusion (IPE) focusing on the involvement of heparanase.. Samples of 30 patients with MPE and 44 patients with IPE were evaluated in comparison to those of 33 patients with transudate pleural effusions, using heparanase ELISA, heparanase procoagulant activity assay, thrombin and factor Xa chromogenic assays and thromboelastography. A cell proliferation assay was performed. EMT-6 breast cancer cells were injected to the pleural cavity of mice. A peptide inhibiting heparanase activity was administered subcutaneously.. Levels of heparanase, factor Xa and thrombin were significantly higher in exudate than transudate. Thromboelastography detected almost no thrombus formation in the whole blood, mainly on MPE addition. This effect was completely reversed by bacterial heparinase. Direct measurement revealed high levels of HS chains in pleural effusions. Higher proliferation was observed in tumour cell lines incubated with exudate than with transudate and it was reduced when bacterial heparinase was added. The tumour size in the pleural cavity of mice treated with the heparanase inhibitor were significantly smaller compared with control (p=0.005).. HS chains released by heparanase form an anticoagulant milieu in MPE, preventing local thrombosis and enabling tumour cell proliferation. Inhibition of heparanase might provide a therapeutic option for patients with recurrent MPE.

Topics: Animals; Anticoagulants; Biomarkers, Tumor; Blood Coagulation; Case-Control Studies; Cell Proliferation; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Glucuronidase; Heparitin Sulfate; Humans; Immunohistochemistry; Male; Mice; Mice, Inbred BALB C; Neovascularization, Pathologic; Pleural Effusion, Malignant; Reference Values; Statistics, Nonparametric; Thrombelastography; Thrombosis; Tumor Cells, Cultured

Syndecan-1 is a proteoglycan that acts as co-receptor through its heparan sulfate (HS) chains and plays important roles in cancer. HS chains are highly variable in length and sulfation pattern. This variability is enhanced by the SULF1/2 enzymes, which remove 6-O-sulfates from HS. We used malignant mesothelioma, an aggressive tumor with poor prognosis, as a model and demonstrated that syndecan-1 over-expression down-regulates SULF1 and alters the HS biosynthetic machinery. Biochemical characterization revealed a 2.7-fold reduction in HS content upon syndecan-1 over-expression, but an overall increase in sulfation. Consistent with low SULF1 levels, trisulfated disaccharides increased 2.5-fold. ERK1/2 activity was enhanced 6-fold. Counteracting ERK activation, Akt, WNK1, and c-Jun were inhibited. The net effect of these changes manifested in G1 cell cycle arrest. Studies of pleural effusions showed that SULF1 levels are lower in pleural malignancies compared to benign conditions and inversely correlate with the amounts of syndecan-1, suggesting important roles for syndecan-1 and SULF1 in malignant mesothelioma.

Topics: Biosynthetic Pathways; Cell Cycle; Cell Line, Tumor; Gene Expression; Gene Expression Regulation, Neoplastic; Heparitin Sulfate; Humans; Kaplan-Meier Estimate; Lung Neoplasms; Mesothelioma; Mesothelioma, Malignant; Pleural Effusion, Malignant; Proportional Hazards Models; Signal Transduction; Sulfotransferases; Syndecan-1