title: PLK-1 Silencing in Bladder Cancer by siRNA Delivered With Exosomes.
abstract: To use exosomes as a vector to deliver small interfering ribonucleic acid (siRNA) to silence the polo-like kinase 1 (PLK-1) gene in bladder cancer cells.
  
  
  
  Exosomes were isolated from both human embryonic kidney 293 (HEK293) cell and mesenchymal stem cell (MSC) conditioned media. Fluorescently labeled exosomes were co-cultured with bladder cancer and normal epithelial cells and uptake was quantified by image cytometry. PLK-1 siRNA and negative control siRNA were loaded into HEK293 and MSC exosomes using electroporation. An invasive bladder cancer cell line (UMUC3) was co-cultured with the electroporated exosomes. Quantitative reverse transcriptase polymerase chain reaction was performed. Protein analysis was performed by Western blot. Annexin V staining and MTT assays were used to investigate effects on apoptosis and viability.
  
  
  
  Bladder cancer cell lines internalize an increased percentage of HEK293 exosomes when compared to normal bladder epithelial cells. Treatment of UMUC3 cells with exosomes electroporated with PLK-1 siRNA achieved successful knockdown of PLK-1 mRNA and protein when compared to cells treated with negative control exosomes.
  
  
  
  HEK293 and MSC exosomes were effectively used as a delivery vector to transport PLK-1 siRNA to bladder cancer cells in vitro, resulting in selective gene silencing of PLK-1. The use of exosomes as a delivery vector for potential intravesical therapy is attractive.
authors.affiliation: Department of Urology, Loyola University Chicago, Maywood, IL.
authors.forename: Kristin A
authors.initials: KA
authors.lastname: Greco
authors.affiliation: Department of Urology, Loyola University Chicago, Maywood, IL.
authors.forename: Carrie A
authors.initials: CA
authors.lastname: Franzen
authors.affiliation: Department of Pathology, Loyola University Chicago, Maywood, IL; Oncology Institute, Loyola University Chicago, Maywood, IL.
authors.forename: Kimberly E
authors.initials: KE
authors.lastname: Foreman
authors.affiliation: Department of Urology, Loyola University Chicago, Maywood, IL.
authors.forename: Robert C
authors.initials: RC
authors.lastname: Flanigan
authors.affiliation: Oncology Institute, Loyola University Chicago, Maywood, IL; Department of Surgery, Loyola University Chicago, Maywood, IL.
authors.forename: Paul C
authors.initials: PC
authors.lastname: Kuo
authors.affiliation: Department of Urology, Loyola University Chicago, Maywood, IL; Oncology Institute, Loyola University Chicago, Maywood, IL; Department of Surgery, Loyola University Chicago, Maywood, IL; Department of Radiology, Loyola University Chicago, Maywood, IL. Electronic address: gogupta@lumc.edu.
authors.forename: Gopal N
authors.initials: GN
authors.lastname: Gupta
mesh_terms: D018797:Cell Cycle Proteins / Q000235:genetics*; D055354:Exosomes; D020868:Gene Silencing; D006801:Humans; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D034741:RNA, Small Interfering; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000235:genetics*; D000097869:Polo-Like Kinase 1
chemical_list: D018797:Cell Cycle Proteins; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; D000097869:Polo-Like Kinase 1