MP34-16 PLK1 SILENCING IN BLADDER CANCER BY SIRNA DELIVERED WITH EXOSOMES

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research III1 Apr 2014MP34-16 PLK1 SILENCING IN BLADDER CANCER BY SIRNA DELIVERED WITH EXOSOMES Kristin A. Greco, Carrie A. Franzen, Paul C. Kuo, Kimberly E. Foreman, Robert C. Flanigan, and Gopal N. Gupta Kristin A. GrecoKristin A. Greco More articles by this author , Carrie A. FranzenCarrie A. Franzen More articles by this author , Paul C. KuoPaul C. Kuo More articles by this author , Kimberly E. ForemanKimberly E. Foreman More articles by this author , Robert C. FlaniganRobert C. Flanigan More articles by this author , and Gopal N. GuptaGopal N. Gupta More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.1029AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The use of small interfering RNAs (siRNAs) in gene therapy is limited by the lack of safe and efficient delivery vectors. Exosomes are nanovesicles that naturally carry RNA and have the ability to induce transcriptional and translational changes in target cells, thus their use as delivery vectors for siRNA is promising. Polo-like kinase-1 (PLK1) gene is a regulator of mitotic progression and is overexpressed in bladder cancer. Studies show that PLK1 depletion leads to cell cycle arrest and apoptosis in bladder cancer cells. Our objective was to use exosomes as a vector to deliver PLK1 siRNA to bladder cancer cells lines. METHODS Exosomes were isolated by ultracentrifugation from human embryonic kidney (HEK) cell conditioned media and labeled with PKH-26 membrane dye. Exosomes were then co-cultured with bladder cancer cells as well as normal bladder epithelial cells and imaged on Amnis ImageStreamX to assess for differential uptake. PLK1 siRNA and negative control siRNA were loaded into HEK exosomes using electroporation. An invasive bladder cancer cell line (UMUC3) was co-cultured with electroporated exosomes. Total RNA was isolated 24 and 48 hours following the addition of exosomes and real-time PCR was performed. RESULTS Bladder cancer cells lines internalize an increased percentage of HEK exosomes when compared to normal bladder epithelial cells (Figure 1). Exosomes electroporated with PLK1 siRNA achieved successful knockdown of PLK1 RNA when compared to the negative control at both 24 and 48 hour timepoints (Figure 2). CONCLUSIONS The higher rate of HEK exosome uptake by cancer cells has implications for using exosomes as a gene delivery vector in the intravesical therapy setting. The exosomes would be preferentially taken up by bladder cancer cells and exert less effect on normal epithelium. HEK exosomes were effectively used as a delivery vector to transport PLK1 siRNA to bladder cancer cells resulting in selective gene silencing of PLK1. The use of exosomes as a delivery vector is attractive as they enter cells easily and have the potential to be non-immunogenic if isolated from an individual patient. © 2014FiguresReferencesRelatedDetailsCited byFranzen C, Blackwell R, Foreman K, Kuo P, Flanigan R and Gupta G (2018) Urinary Exosomes: The Potential for Biomarker Utility, Intercellular Signaling and Therapeutics in Urological MalignancyJournal of Urology, VOL. 195, NO. 5, (1331-1339), Online publication date: 1-May-2016. Volume 191Issue 4SApril 2014Page: e367-e368 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Kristin A. Greco More articles by this author Carrie A. Franzen More articles by this author Paul C. Kuo More articles by this author Kimberly E. Foreman More articles by this author Robert C. Flanigan More articles by this author Gopal N. Gupta More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...