Shafat Itay, Zcharia Eyal, Nisman Benjamin, Nadir Yona, Nakhoul Farid, Vlodavsky Israel, Ilan Neta
Cancer and Vascular Biology Research Center, The Bruce Rappaport Faculty of Medicine, Technion, Haifa 31096, Israel.
Biochem Biophys Res Commun. 2006 Mar 24;341(4):958-63. doi: 10.1016/j.bbrc.2006.01.048. Epub 2006 Jan 24.
Heparanase is a mammalian endo-beta-D-glucuronidase that cleaves heparan sulfate side chains at a limited number of sites. Heparanase enzymatic activity is thought to participate in degradation and remodeling of the extracellular matrix and to facilitate cell invasion associated with tumor metastasis, angiogenesis, and inflammation. Traditionally, heparanase activity was well correlated with the metastatic potential of a large number of tumor-derived cell types. More recently, heparanase upregulation was detected in an increasing number of primary human tumors, correlating, in some cases, with poor postoperative survival and increased tumor vascularity. The present study was undertaken to develop a highly sensitive ELISA suitable for the determination and quantification of human heparanase in tissue extracts and body fluids. The assay preferentially detects the 8+50 kDa active heparanase heterodimer vs. the latent 65 kDa proenzyme and correlates with immunoblot analysis of heparanase containing samples. It detects heparanase at concentrations as low as 200 pg/ml and is suitable for quantification of heparanase in tissue extracts and urine.
乙酰肝素酶是一种哺乳动物内切β-D-葡糖醛酸酶，可在有限数量的位点切割硫酸乙酰肝素侧链。乙酰肝素酶的酶活性被认为参与细胞外基质的降解和重塑，并促进与肿瘤转移、血管生成和炎症相关的细胞侵袭。传统上，乙酰肝素酶活性与大量肿瘤衍生细胞类型的转移潜能密切相关。最近，在越来越多的原发性人类肿瘤中检测到乙酰肝素酶上调，在某些情况下，这与术后生存率低和肿瘤血管生成增加相关。本研究旨在开发一种高度敏感的酶联免疫吸附测定法（ELISA），适用于测定和定量组织提取物和体液中的人乙酰肝素酶。该测定法优先检测8 + 50 kDa活性乙酰肝素酶异二聚体，而非潜在的65 kDa酶原，并且与含乙酰肝素酶样品的免疫印迹分析相关。它能检测低至200 pg/ml浓度的乙酰肝素酶，适用于定量组织提取物和尿液中的乙酰肝素酶。
