Increased chemotactic migration and growth in heparanase-overexpressing human U251n glioma cells

Department of Neurology, Henry Ford Health Science Center, Detroit, MI, USA.
Journal of Experimental & Clinical Cancer Research (Impact Factor: 3.27). 07/2008; 27(1):23. DOI: 10.1186/1756-9966-27-23
Source: PubMed

ABSTRACT Heparanase is an endoglycosidase that degrades heparan sulfate, the main polysaccharide constituent of the extracellular matrix (ECM) and basement membrane. Expression of the heparanase gene is associated with the invasion and metastatic potential of a variety of tumor-derived cell types. However, the roles of heparanase in the regulation of gene expression and the subsequent cell function changes other than invasion are not clear. In the current study, we overexpressed the human heparanase gene in a human U251n glioma cell line. We found that heparanase-overexpression significantly increased cell invasion, proliferation, anchorage-independent colony formation and chemotactic migration towards fetal bovine serum (FBS)-supplied medium and stromal cell-derived factor-1 (SDF-1). These phenotypic appearances were accompanied by enhanced protein kinase B (AKT) phosphorylation. Focal adhesion kinase (FAK) and extracellular signal-regulated kinase 1 (ERK1) signaling were not altered by heparanase-overexpression. These results indicate that heparanase has pleiotropic effects on tumor cells.

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    FEBS Journal 10/2014; 281(22). DOI:10.1111/febs.13097 · 3.99 Impact Factor
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    ABSTRACT: BACKGROUND: Cell adhesion molecules (CAMs) are expressed ubiquitously. Each of the four families of CAMs is comprised of glycosylated, membrane-bound proteins that participate in multiple cellular processes including cell-cell communication, cell motility, inside-out and outside-in signaling, tumorigenesis, angiogenesis and metastasis. Intercellular adhesion molecule-2 (ICAM-2), a member of the immunoglobulin superfamily of CAMs, has six N-linked glycosylation sites at amino acids (asparagines) 47, 82, 105, 153, 178 and 187. Recently, we demonstrated a previously unknown function for ICAM-2 in tumor cells. We showed that ICAM-2 suppressed neuroblastoma cell motility and growth in soft agar, and induced a juxtamembrane distribution of F-actin in vitro. We also showed that ICAM-2 completely suppressed development of disseminated tumors in vivo in a murine model of metastatic NB. These effects of ICAM-2 on NB cell phenotype in vitro and in vivo depended on the interaction of ICAM-2 with the cytoskeletal linker protein alpha-actinin. Interestingly, ICAM-2 did not suppress subcutaneous growth of tumors in mice, suggesting that ICAM-2 affects the metastatic but not the tumorigenic potential of NB cells. The goal of the study presented here was to determine if the glycosylation status of ICAM-2 influenced its function in neuroblastoma cells. METHODS: Because it is well documented that glycosylation facilitates essential steps in tumor progression and metastasis, we investigated whether the glycosylation status of ICAM-2 affected the phenotype of NB cells. We used site-directed mutagenesis to express hypo- or non-glycosylated variants of ICAM-2, by substituting alanine for asparagine at glycosylation sites, and compared the impact of each variant on NB cell motility, anchorage-independent growth, interaction with intracellular proteins, effect on F-actin distribution and metastatic potential in vivo. RESULTS: The in vitro and in vivo phenotypes of cells expressing glycosylation site variants differed from cells expressing fully-glycosylated ICAM-2 or no ICAM-2. Most striking was the finding that mice injected intravenously with NB cells expressing glycosylation site variants survived longer (P <= 0.002) than mice receiving SK-N-AS cells with undetectable ICAM-2. However, unlike fully-glycosylated ICAM-2, glycosylation site variants did not completely suppress disseminated tumor development. CONCLUSIONS: Reduced glycosylation of ICAM-2 significantly attenuated, but did not abolish, its ability to suppress metastatic properties of NB cells.
    BMC Cancer 05/2013; 13(1):261. DOI:10.1186/1471-2407-13-261 · 3.32 Impact Factor
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    ABSTRACT: Primary cell culture is a valuable tool that extensively used in the basic research of organ specific functions, physiology, pathology, pharmacology and so on. The aim of this study was to derive primary mesenchyme cell line representative of colon tissues. First, we identified the location of sub-epithelial myofibroblast in the colon tissues in rat. Alpha-smooth muscle-actin (alpha-SMA) positive myofibroblast found just beneath the epithelia of colon tissues. Accordingly, we isolated intestinal sub-epithelial myofibroblasts (ISEMFs) from colonic lamina propria of rat. The isolated cells were cultured and characterized by phase contrast, immunofluorescent and RT-PCR analysis. The harvested cells expressed the specific markers such as alpha-SMA, vimentin and were negative for desmin expression. Finally, we did trans-well migration assay to know the difficulties or convenience of this isolated cell culture experiment. The isolated rat ISEMFs grow well in culture media and easy to culture for the research purposes. Therefore, the primary ISEMF might be a useful alternative in vitro tool for bioresearch study.

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