154 KNOCKDOWN OF INTEGRIN-LINKED KINASE REDUCES INVASIVE AND METASTATIC POTENTIAL OF RENAL CELL CARCINOMA

Abstract

You have accessJournal of UrologyKidney Cancer: Basic Research I1 Apr 2012154 KNOCKDOWN OF INTEGRIN-LINKED KINASE REDUCES INVASIVE AND METASTATIC POTENTIAL OF RENAL CELL CARCINOMA Kyung Seok Han, Peter Raven, Shannon Awrey, Estelle Li, Ladan Fazli, Martin Gleave, and Alan So Kyung Seok HanKyung Seok Han Vancouver, Canada More articles by this author , Peter RavenPeter Raven Vancouver, Canada More articles by this author , Shannon AwreyShannon Awrey Vancouver, Canada More articles by this author , Estelle LiEstelle Li Vancouver, Canada More articles by this author , Ladan FazliLadan Fazli Vancouver, Canada More articles by this author , Martin GleaveMartin Gleave Vancouver, Canada More articles by this author , and Alan SoAlan So Vancouver, Canada More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.204AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Integrin-linked kinase (ILK) is a serine/threonine kinase implicated in the regulation of cell growth and survival, cell cycle progression, epithelial-mesenchymal transition (EMT), invasion and migration, angiogenesis. The role of ILK has not been evaluated in renal cell carcinoma (RCC). We investigated the role of ILK on cancer progression and metastasis and therapeutic potential of ILK inhibition in RCC. METHODS Baseline expression of ILK was evaluated by immunofluorescence and western blotting in non-cancerous renal tubular cells (HK-2) and RCC cells (UMRC-6, UMRC-3 and Caki-1). Molecular EMT markers were examined by Western blotting. RNAi using siRNA was used to knock down ILK in vitro. After transient transfection, crystal violet assay and cell cycle analysis using FACS were performed to assess the effect of ILK on tumor growth and cell cycle. Confocal microscopy was used to evaluate changes in stress fibers and focal adhesions with phalloidin-rhodamine and vinculin antibody. Scratch assay and Matrigel invasion assay were performed to evaluate changes of EMT phenotypes after knockdown of ILK. RESULTS ILK is relatively less expressed in normal cells (HK-2) and low stage RCC cells (UMRC-6) but highly expressed in advanced and metastatic RCC cells (UMRC-3 and Caki-1). Caki-1, metastatic RCC cells showed higher expressions of molecular EMT markers including Snail, Zeb1 but decreased activity of GSK3β. Knockdown of ILK using si-ILK inhibited tumor proliferation but the inhibition rate was less than 10% and cell cycle progression was not significantly affected by ILK inhibition. However, ILK knockdown suppressed formation of stress fibers and focal adhesions in UMRC-3 and Caki-1 cells and also effectively suppressed phenotypic EMT markers including cell migration and invasion in Caki-1 and UMRC-3 cells. CONCLUSIONS ILK is highly expressed in advanced RCC and its high expression is related to EMT-related protein in RCC. Knockdown of ILK inhibited molecular EMT markers and suppressed cell migration and invasion. These results suggest the therapeutic potential of ILK inhibition on invasion and metastasis in advanced RCC. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e63-e64 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Kyung Seok Han Vancouver, Canada More articles by this author Peter Raven Vancouver, Canada More articles by this author Shannon Awrey Vancouver, Canada More articles by this author Estelle Li Vancouver, Canada More articles by this author Ladan Fazli Vancouver, Canada More articles by this author Martin Gleave Vancouver, Canada More articles by this author Alan So Vancouver, Canada More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...