MP65-05 NOTCH2 IN THE BLADDER PROMOTES TUMOR DEVELOPMENT AND LEADS TO MORE MALIGNANT PHENOTYPE.

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology III1 Apr 2018MP65-05 NOTCH2 IN THE BLADDER PROMOTES TUMOR DEVELOPMENT AND LEADS TO MORE MALIGNANT PHENOTYPE. Akihiro Goriki, Takeshi Sano, Alberto Contreras, Htoo Oo, Tetsutaro Hayashi, Akio Matsubara, and Peter Black Akihiro GorikiAkihiro Goriki More articles by this author , Takeshi SanoTakeshi Sano More articles by this author , Alberto ContrerasAlberto Contreras More articles by this author , Htoo OoHtoo Oo More articles by this author , Tetsutaro HayashiTetsutaro Hayashi More articles by this author , Akio MatsubaraAkio Matsubara More articles by this author , and Peter BlackPeter Black More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.2071AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Our recently published results revealed that NOTCH2 is an oncogene that drives bladder cancer (BCa) progression. To test whether NOTCH2 can promote BCa development, we have established a constitutively active NOTCH2 intracellular domain (N2ICD) mouse model. Previous TCGA data showed that NOTCH2 expression and copy number gain are enriched in basal tumors. METHODS We have created lentiviral constructs based on the FUGW vector containing the N2ICD transgene, driven by either uroplakin-2 (Upk2, to induce luminal tumors) or cytokeratin-5 (Krt5, to induce basal tumors). These constructs also include intraribosomal entry sequence (IRES) followed by the firefly luciferase gene. Lentiviral particles were inocluted by ultra-sound guided injections into the subepithelial space of the bladder wall. Mice were treated with the bladder specific carcinogen BBN (0.05%, in drinking water) for 12 weeks. We have also tested the metastatic potential of NOTCH2 mutants (A2025E and Q2223X) over-expressed cell lines in vivo using the zebrafish embryo model. Each wild-type, NOTCH2 mutants (A2025E and Q2223X), N2ICD over-expressed RT4 or RT112 cell lines, and NOTCH2 knock-down UC13 cell lines were injected to zebrafish embryo. After 3 days, the number of metastases were counted. RESULTS Luciferase activity was observed in the bladder of some mice 20 weeks into the experiment: 2 cases in the empty vector control group (2/15, 13.3%), 6 cases in the Upk2 group (6/15, 40%), and 5 cases in Krt5 group (5/15, 33.3%). The tumors in both Upk2 and Krt5 group showed not only urothelial but also squamous cell carcinoma features. Moreover, in both Upk2 and Krt5 group, tumorigenesis was observed in 20 weeks after BBN while tumorigenesis in control group was observed in 26 weeks. In the zebrafish model, the number of metastasis were increased in the N2ICD over-expressed groups compared to the wild-type and mutant lines, while the metastasis were decreased in NOTCH2 knock-down UC13 line. CONCLUSIONS Our results suggest that the over-expression of N2ICD in the bladder wall could accelerate tumor development and possibly lead to more malignant phenotypes. Moreover, over-expression of N2ICD promotes metastatic potential. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e860 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Akihiro Goriki More articles by this author Takeshi Sano More articles by this author Alberto Contreras More articles by this author Htoo Oo More articles by this author Tetsutaro Hayashi More articles by this author Akio Matsubara More articles by this author Peter Black More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...