Abstract
Abstract OBJECTIVE: The aim of this study was to evaluate the role of membrane protein urokinase-type plasminogen activator receptor associated protein (uPARAP), which is one of the member of serine-protease cascade, urokinase plasminogen activator system, in the motile and invasive activities of glioma cells. METHODS: In the present study, we first evaluate expression of uPARAP in various normal tissues as well as glioma patients’ samples and glioma cell lines using quantative PCR and Western blotting. To evaluate the function of uPARAP, we silence the uPARAP gene using two independent siRNAs designed for uPARAP, and evaluate its function on motile and invasive activities of glioma cells by Boyden chamber migration and invasion assays. We also evaluate the function of uPARAP on rearrangement of actin cytoskeleton by staining glioma cells with Alexa Fluor 488 conjugated Phalloidin. RESULTS: uPARAP protein was expressed four out of four (100%) glioma samples regardless of its World Health Organization grade but did not express in normal brain control. Introduction of small interfering RNA targeted uPARAP into two different glioma cell lines; KNS42 and KNS81, resulted in down-reguration of uPARAP expression, and significantly suppressed glioma cells’ migration and invasion properties in vitro. Rearrangement of actin cytoskeleton was observed at Alexa Fluor 488 conjugated Phalloidin staining. In uPARAP knocked-down glioma cells, polymeric actin became organized in stress fibers and the lamellipodia disappeared. CONCLUSION: On the basis of our findings, we conclude that RNA interference-mediated down-regulation of uPARAP decreases invasion and migration property in glioma cells. The inhibition of invasion and migration property was mediated via reorganization of the actin cytoskeleton. Downregulation of uPARAP may be a novel anti-invasion therapeutic strategy for malignant gliomas. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 531.
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