Document Detail

Colorimetric heparinase assay for alternative anti-metastatic activity.
MedLine Citation:
PMID:  16806278     Owner:  NLM     Status:  MEDLINE    
Heparanase has been previously associated with the metastatic potential, inflammation, and angiogenesis of tumor cells. Heparanase activity has been detected by means of UV absorption, radiolabeled substrates, electrophoretic migration, and heparan sulfate affinity assays. However, those methods have proven to be somewhat problematic with regards to application to actual biological samples, the accessibility of the immobilized substrates, experimental sensitivity, and the separation of degraded products. Rather than focusing on heparanase activity, then, we have developed a rapid, alternative colorimetric heparinase assay, on the basis of the recent finding that sulfated disaccharides generated from heparin by bacterial heparinase exhibit biological properties comparable to those from heparan sulfate by mammalian heparanase. In this study, the concentrations of porcine heparin and bacterial heparinase I were determined using a Sigma Diagnostics Kit. Morus alba was selected as a candidate through this assay system, and an inhibitor, resveratrol, was purified from its methanol extract. Its anti-metastatic effects on the pulmonary metastasis of murine B16 melanoma cells were also evaluated. Our findings suggest that this assay may prove useful as a diagnostic tool for heparinase inhibition, as an alternative anti-metastatic target.
S C Ahn; B Y Kim; W K Oh; Y M Park; H M Kim; J S Ahn
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2006-05-27
Journal Detail:
Title:  Life sciences     Volume:  79     ISSN:  0024-3205     ISO Abbreviation:  Life Sci.     Publication Date:  2006 Sep 
Date Detail:
Created Date:  2006-09-18     Completed Date:  2006-11-02     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0375521     Medline TA:  Life Sci     Country:  England    
Other Details:
Languages:  eng     Pagination:  1661-5     Citation Subset:  IM    
Department of Microbiology and Immunology, Pusan National University College of Medicine, Ami-dong 1-10, Seo-ku, Pusan 602-739, Korea.
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MeSH Terms
Antineoplastic Agents / pharmacology*
Biological Assay
Cell Line, Tumor
Colorimetry / methods
Drug Screening Assays, Antitumor
Enzyme Inhibitors / analysis,  pharmacology*
Flavobacterium / enzymology
Heparin / analysis,  metabolism
Heparin Lyase / analysis*,  antagonists & inhibitors
Intestinal Mucosa
Lung Neoplasms / drug therapy,  pathology,  secondary
Melanoma / drug therapy,  pathology,  secondary
Mice, Inbred C57BL
Morus / chemistry
Neoplasm Metastasis / drug therapy*,  pathology
Plant Extracts / pharmacology*
Specific Pathogen-Free Organisms
Reg. No./Substance:
0/Antineoplastic Agents; 0/Enzyme Inhibitors; 0/Plant Extracts; 9005-49-6/Heparin; EC Lyase

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