MP92-07 NETRIN-1: A NOVEL PROTEIN IDENTIFIED IN DISEASE PROGRESSION OF SUNITINIB RESISTANT RENAL CELL CARCINOMA

Abstract

You have accessJournal of UrologyKidney Cancer: Basic Research & Pathophysiology III1 Apr 2016MP92-07 NETRIN-1: A NOVEL PROTEIN IDENTIFIED IN DISEASE PROGRESSION OF SUNITINIB RESISTANT RENAL CELL CARCINOMA Sebastian Frees, Claudia Chavez-Munoz, Betty Zhou, Alexander Wong, Peter Raven, and Alan So Sebastian FreesSebastian Frees More articles by this author , Claudia Chavez-MunozClaudia Chavez-Munoz More articles by this author , Betty ZhouBetty Zhou More articles by this author , Alexander WongAlexander Wong More articles by this author , Peter RavenPeter Raven More articles by this author , and Alan SoAlan So More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.2640AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Clear-cell renal cell carcinoma (ccRCC) is the sixth most common malignancy in men in North America with an incidence that has been steadily increasing at a rate of 3% yearly over the last three decades. Patients with ccRCC can be treated with surgery; however, 30% of first-time diagnosed patients present with metastatic disease. Since ccRCC is a malignancy dependent on neovascularization, current first line systemic therapies target the formation of new vessels allowing nutrient depravation and cell death. A family of drugs that uses this mechanism is called Tyrosine kinase inhibitors (TKI), and one of the members of this family is Sunitinib. Sunitinib is the first-line of therapy currently used in patients with ccRCC. However, recent studies have shown that after approximately 1 year of treatment patients develop resistance, showing disease progression while on therapy. Therefore in this study we propose to identify the protein(s) responsible for increase migration and invasion with the aim of developing a new therapy that will target the identified protein and potentially slow down the progression of the disease. METHODS Human renal cancer cell lines were used (CAKI-1, CAKI-2, ACHN) and treated with Sunitinib at increasing doses to develop a Sunitinib conditioned renal cell carcinoma cell line. A mRNA microarray was performed to compare the differences in gene expression between CAKI-1 Sunitinib conditioned cell line and CAKI-1 non-conditioned cell line. Moreover, we have successfully established an orthotopic renal cell carcinoma Sunitinib resistant animal model. Western blots and q-PCR were used to confirm microarray results and to evaluate the expression of Netrin-1 in-vitro. XCELLigence system was used to evaluate migration and invasion of CAKI-1 Sunitinib conditioned cell lines. Immunostaining of our in vivo Sunitinib resistant model confirms the in vitro findings. RESULTS Human renal cell carcinoma Sunitinib conditioned cell lines showed a highly upregulated expression of Netrin-1 in the microarray results as well as in Western blotting and q-PCR when compared to the un-treated renal cell carcinoma cell lines. XCELLigence system demonstrated an increased migration in CAKI-1 Sunitinib conditioned cell lines when compared to the non-treated ones as well as, increased endothelial cell migration when co-cultured with CAKI-1 Sunitinib conditioned cell line. Our results also demonstrated an increase expression of UNC5B and DCC Netrin-1 receptors in CAKI-1 Sunitinib conditioned cell line. Silencing of Netrin-1 in CAKI-1 Sunitinib conditioned cell line demonstrated a significant reduction in cell migration and invasion. Interestingly, our orthotopic renal cell carcinoma Sunitinib resistance animal model as well as human renal cell carcinoma Sunitinib resistant tumors confirmed a high expression of Netrin-1 when compared to those of control. CONCLUSIONS In conclusion we have found that Netrin-1 plays a key role in the migration and progression of renal cell carcinoma. Development of therapies targeting Netrin-1 suggests to be a promising approach to suppress renal cell carcinoma tumor progression and potentially prolong patients' survival. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e1164 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Sebastian Frees More articles by this author Claudia Chavez-Munoz More articles by this author Betty Zhou More articles by this author Alexander Wong More articles by this author Peter Raven More articles by this author Alan So More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...