46 These early observations gained substantial support when specific molecular probes became available shortly after cloning of the Onalespib clinical trial heparanase gene. Both over-expression and silencing (Figure 3) of the heparanase gene clearly indicate that heparanase not only enhances cell dissemination but also promotes the establishment of a vascular network that accelerates primary tumor growth and provides a gateway for invading metastatic cells.16 While these studies provided a proof-of-concept for the prometastatic and proangiogenic

capacity of heparanase, the clinical significance of the enzyme Inhibitors,research,lifescience,medical in tumor progression emerged from a systematic evaluation of heparanase expression in primary human tumors. Heparanase has been found to be up-regulated in essentially all human carcinomas and Inhibitors,research,lifescience,medical sarcomas examined.16 Notably, increased heparanase levels were most often associated with reduced patient survival post operation, increased tumor metastasis, and higher microvessel density.16,47 Figure 3 Lung colonization of B16 mouse melanoma cells is inhibited following silencing (sM2 antiheparanase siRNA) of the heparanase gene. Both gene expression (A: RT-PCR)

and lung metastasis (B, C) are inhibited by 80%–90% upon silencing of the endogenous … The Inhibitors,research,lifescience,medical cellular and molecular mechanisms underlying enhanced tumor growth by heparanase are only starting to be revealed. At the cellular level, both tumor cells and cells that comprise the tumor microenvironment (i.e. endothelial, fibroblasts, tumor-infiltrating Inhibitors,research,lifescience,medical immune cells) are likely to be affected by heparanase. Proangiogenic potency of heparanase was established clinically16,48 and in several in-vitro and in-vivo model systems,

including wound-healing,49,50 tumor xenografts,51 Matrigel plug assay,49 and tube-like structure formation. Moreover, microvessel density was significantly reduced in tumor xenografts developed by T lymphoma cells transfected Inhibitors,research,lifescience,medical with antiheparanase ribozyme.52 The molecular mechanism by which heparanase facilitates angiogenic responses has traditionally been attributed primarily to the release of HS-bound growth factors such as VEGF-A and FGF-2,18,53 a direct consequence of heparanase enzymatic activity. Heparanase was also noted to facilitate Ergoloid the formation of lymphatic vessels. In head and neck carcinoma, high levels of heparanase were associated with increased lymphatic vessel density (LVD), increased tumor cell invasion to lymphatic vessels, and increased expression of VEGF-C,54 a potent mediator of lymphatic vessel formation. Heparanase over-expression by melanoma, epidermoid, breast and prostate carcinoma cells induced a 3–5-fold elevation of VEGF-C expression in vitro, and facilitated lymph angiogenesis of tumor xenografts in vivo, whereas heparanase gene silencing was associated with decreased VEGF-C levels.