CDK4/6 Inhibition Controls Proliferation of Bladder Cancer and Transcription of RB1

No AccessJournal of UrologyInvestigative Urology1 Mar 2016CDK4/6 Inhibition Controls Proliferation of Bladder Cancer and Transcription of RB1 Anuja Sathe, Nicole Koshy, Sebastian C. Schmid, Mark Thalgott, Sarah M. Schwarzenböck, Bernd J. Krause, Per S. Holm, Juergen E. Gschwend, Margitta Retz, and Roman Nawroth Anuja SatheAnuja Sathe More articles by this author , Nicole KoshyNicole Koshy More articles by this author , Sebastian C. SchmidSebastian C. Schmid More articles by this author , Mark ThalgottMark Thalgott More articles by this author , Sarah M. SchwarzenböckSarah M. Schwarzenböck More articles by this author , Bernd J. KrauseBernd J. Krause More articles by this author , Per S. HolmPer S. Holm More articles by this author , Juergen E. GschwendJuergen E. Gschwend More articles by this author , Margitta RetzMargitta Retz More articles by this author , and Roman NawrothRoman Nawroth More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.08.082AboutFull TextPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract Purpose: The retinoblastoma signaling network is frequently altered in advanced bladder cancer. We investigated the potential of CDK4/6 as a therapeutic target and determined biomarkers for patient stratification. Materials and Methods: Genetic alterations were analyzed using public databases, including TCGA (The Cancer Genome Atlas), COSMIC (Catalogue of Somatic Mutations in Cancer) and CCLE (Cancer Cell Line Encyclopedia). Effects of the CDK4/6-inhibitor PD-0332991 or LY2835219 were examined in 10 bladder cancer cell lines by immunoblot, cell viability, apoptosis and cell cycle progression. Efficacy of the PD-0332991 and cisplatin combination was analyzed using the combination index. Gene expression level was determined by quantitative polymerase chain reaction. Cytomegalovirus promoter regulated recombinant retinoblastoma was used for reconstitution. Three-dimensional xenografts were grown on chicken chorioallantoic membrane and analyzed by measuring tumor weight and immunohistochemical expression of total retinoblastoma and Ki-67. Results: PD-0332991 treatment decreased the proliferation of retinoblastoma positive bladder cancer cell lines and was synergistic in combination with cisplatin. PD-0332991 or LY2835219 treatment decreased the phosphorylation, total protein and transcript level of retinoblastoma. Treatment resulted in a decrease in E2F target gene expression (CCNA2 and CCNE2) and cell cycle progression from G0/G1 to the S-phase but did not affect apoptosis. In retinoblastoma negative cells reconstituted with recombinant retinoblastoma PD-0332991 affected only phosphorylation and not the total retinoblastoma level. These cells remained resistant to treatment. In 3-dimensional retinoblastoma xenografts, treatment resulted in reduced tumor weight and decreased expression of total retinoblastoma and Ki-67. Conclusions: We provide preclinical evidence that CDK4/6 inhibition is a potential therapeutic strategy for retinoblastoma positive bladder cancer that probably acts by negatively regulating retinoblastoma transcription. References 1 : Cancer statistics, 2015. CA Cancer J Clin2015; 65: 5. Google Scholar 2 : EAU guidelines on muscle-invasive and metastatic bladder cancer: summary of the 2013 guidelines. Eur Urol2014; 65: 778. Google Scholar 3 : Long-term survival results of a randomized trial comparing gemcitabine plus cisplatin, with methotrexate, vinblastine, doxorubicin, plus cisplatin in patients with bladder cancer. 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Bob Weinberg provided RcCMV/Rb.MetricsAuthor Information Anuja Sathe More articles by this author Nicole Koshy More articles by this author Sebastian C. Schmid More articles by this author Mark Thalgott More articles by this author Sarah M. Schwarzenböck More articles by this author Bernd J. Krause More articles by this author Per S. Holm More articles by this author Juergen E. Gschwend More articles by this author Margitta Retz More articles by this author Roman Nawroth More articles by this author Expand All Advertisement PDF downloadLoading ...