587 LOSS OF BCG VIABILITY ADVERSELY AFFECTS THE DIRECT RESPONSE OF UROTHELIAL CARCINOMA CELLS TO BCG EXPOSURE

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research (I)1 Apr 2013587 LOSS OF BCG VIABILITY ADVERSELY AFFECTS THE DIRECT RESPONSE OF UROTHELIAL CARCINOMA CELLS TO BCG EXPOSURE Guangjian Zhang, Fanghong Chen, Yanli Cao, Jacek Zielonka, Gang Cheng, Gopit Shah, Balaraman Kalyanaraman, and William See Guangjian ZhangGuangjian Zhang Milwaukee, WI More articles by this author , Fanghong ChenFanghong Chen Milwaukee, WI More articles by this author , Yanli CaoYanli Cao Milwaukee, WI More articles by this author , Jacek ZielonkaJacek Zielonka Milwaukee, WI More articles by this author , Gang ChengGang Cheng Milwaukee, WI More articles by this author , Gopit ShahGopit Shah Milwaukee, WI More articles by this author , Balaraman KalyanaramanBalaraman Kalyanaraman Milwaukee, WI More articles by this author , and William SeeWilliam See Milwaukee, WI More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.1983AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Previous studies have demonstrated that in both the laboratory and clinical setting BCG viability is a variable that correlates with anti-tumor efficacy. Our group has demonstrated that loss of BCG viability reduces BCG induced cellular necrosis and HMGB1 release, a protein that is required for the in vivo anti-tumor effect of BCG. This study evaluated how loss of BCG viability impacts additional molecular and phenotypic intermediate endpoints that characterize, and/or contribute to, the direct effect of BCG on UC cells. METHODS Two human UC cell lines (253J amd T24) were used to study the effect of loss of BCG viability on the tumor cell response to BCG. The cellular response to BCG rendered non-viable by heat killing (hk) was compared to the response to viable BCG. The response endpoints evaluated included the induction of oxidative stress (iNOS expression), activation of intracellular signaling pathways (NFκB and NRF2), gene transactivation (IL6, IL8, CXCL1, CXCL3, CCL20, and CD54), and cytotoxicity (membrane integrity, LDH release, MTT). RESULTS Loss of viability (hkBCG) resulted in a quantitative decrease in the tumor response, relative to viable BCG, for all of the measured endpoints. The magnitude and statistical significance of the decrease in response varied by cell line and endpoint, ranging from 15% to 100% of the response to viable BCG. The attached table provides average fold changes for specific endpoints, relative to control cells,for the viable and hkBCG treatment groups. While quantitatively different non-viable BCG continued to induce responses that were qualitatively similar to BCG relative to untreated controls. CONCLUSIONS BCG viability is an important variable influencing the direct tumor cell response to BCG. Loss of BCG viability results in quantitative reductions in the magnitude of BCG's direct effects on UC cell biology. However non-viable BCG continues to have activity across the range of biologic endpoints evaluated in this study. Further study will be required to identify the underlying mechanism through which BCG's activity is attenuated as a consequence of loss of viability. Average fold change in tumor cell response endpoint for cells exposed to either viable or heat-killed BCG, relative to controls. iNOS NRF2 NFkB p21 Gene Expression LDH Release Vital Dye Exclusion Viable BCG 6.6 2.0 2.1 2.1 15 3.8 13 Heat Killed BCG 4.5 1.5 1.8 1.7 6 3.6 6 P Value (viable vs HK) = 0.15 < 0.01 = 0.3 < 0.01 < 0.05 = 0.3 < 0.001 © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e240 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Guangjian Zhang Milwaukee, WI More articles by this author Fanghong Chen Milwaukee, WI More articles by this author Yanli Cao Milwaukee, WI More articles by this author Jacek Zielonka Milwaukee, WI More articles by this author Gang Cheng Milwaukee, WI More articles by this author Gopit Shah Milwaukee, WI More articles by this author Balaraman Kalyanaraman Milwaukee, WI More articles by this author William See Milwaukee, WI More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...