306 ROLE OF EPHA2 IN MALIGNANT CELLULAR BEHAVIOR IN RENAL CELL CARCINOMA CELLS

Abstract

You have accessJournal of UrologyKidney Cancer: Basic Research (II)1 Apr 2013306 ROLE OF EPHA2 IN MALIGNANT CELLULAR BEHAVIOR IN RENAL CELL CARCINOMA CELLS Min Chul Cho, Sung Yong Cho, Seung Bae Lee, Cheol Kwak, Hae Won Lee, Hyeon Hoe Kim, and Hyeon Jeong Min Chul ChoMin Chul Cho Goyang, Korea, Republic of More articles by this author , Sung Yong ChoSung Yong Cho Seoul, Korea, Republic of More articles by this author , Seung Bae LeeSeung Bae Lee Seoul, Korea, Republic of More articles by this author , Cheol KwakCheol Kwak Seoul, Korea, Republic of More articles by this author , Hae Won LeeHae Won Lee Goyang, Korea, Republic of More articles by this author , Hyeon Hoe KimHyeon Hoe Kim Seoul, Korea, Republic of More articles by this author , and Hyeon JeongHyeon Jeong Seoul, Korea, Republic of More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.1690AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Increased EphA2 expression can promote tumor progression by inducing cell growth and invasion, while concurrently decreasing apoptosis in the tumor cells of various types of cancer. Recently, it has been shown that focal adhesion kinase (FAK) or RhoA can act as an important signaling molecule that mediates EphA2 activation to cellular responses such as migration, invasion and suppression of apoptosis in several types of cancer. To date, there have been no studies which have investigated EphA2 expression or its role in malignant cellular behavior or the relationships among EphA2, FAK, and RhoA on renal cell carcinoma (RCC) cells. We investigated the role of EphA2 in malignant cellular behavior on RCC cells and whether phosphorylated FAK and active RhoA can act as downstream effectors of EphA2 on RCC cells. METHODS Expression of EphA2 protein in non-metastatic RCC (Caki-2 and A498), metastatic RCC cells (Caki-1 and ACHN), human embryonic kidney-293 (HEK-293) cell and human prostate cancer cell lines (PC-3 and DU-145) was evaluated by Western blot. PC-3 and DU-145 were used as positive controls of EphA2 expression. Changes in expression of EphA2 following transfection with EphA2 small interfering RNA (siRNA) were determined by reverse transcription polymerase chain reaction and Western blot. Cellular viability, apoptosis and invasion were analyzed by cell counting kit-8, Annexin-V and modified Matrigel-Boyden assays, respectively. The effect of EphA2 siRNA on phosphorylation of FAK and expression of activated RhoA protein were evaluated by Western blot. RESULTS In all RCC cell lines, the expression of EphA2 protein was detectable at variable levels while that in HEK-293 cell was very low. In all RCC cell lines, the expression of EphA2 mRNA and protein at 48-hours after EphA2 siRNA transfection was significantly lower than that after transfection with control siRNA or that in untreated cells. In the non-metastatic RCC cells (Caki-2 and A498), cellular viability, invasiveness, resistance to apoptosis, the expression of active RhoA protein and FAK phosphorylation were significantly decreased in EphA2 siRNA-treated cells compared to cells treated with control siRNA or untreated control, while not in the metastatic RCC cells (Caki-1 or ACHN). CONCLUSIONS Our data provide the first functional evidence that EphA2 may play a critical role in malignant cellular behavior of RCC such as invasion and resistance to apoptosis and appears to be functional particularly in the early stage of malignant progression of non-metastatic RCC. Also, FAK and RhoA might act as downstream effectors of EphA2. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e124 Peer Review Report Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Min Chul Cho Goyang, Korea, Republic of More articles by this author Sung Yong Cho Seoul, Korea, Republic of More articles by this author Seung Bae Lee Seoul, Korea, Republic of More articles by this author Cheol Kwak Seoul, Korea, Republic of More articles by this author Hae Won Lee Goyang, Korea, Republic of More articles by this author Hyeon Hoe Kim Seoul, Korea, Republic of More articles by this author Hyeon Jeong Seoul, Korea, Republic of More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...