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iNOS expression and NO production contribute to the direct effects of BCG on urothelial carcinoma cell biology. Urol Oncol 2014 Jan;32(1):45.e1-9 PMID: 24054867 PMCID: PMC5798243

Pubmed ID





OBJECTIVES: Evidence suggests that oxidative stress occurring as a consequence of inducible nitric oxide synthase/nitric oxide (iNOS/NO) contributes to the biologic effects of bacille Calmette-Guérin (BCG). Objective of this study is to examine iNOS expression, NO production, and the biologic effect of NO on established intermediate end points for the human urothelial carcinoma cell response to BCG.

MATERIALS AND METHODS: Quantitative reverse transcriptase-polymerase chain reaction and real-time measurement of NO was used to assess iNOS and NO production, respectively, in 2 human urothelial carcinoma (UC) cell lines, in response to BCG. The effect of blocking NO production using the specific iNOS inhibitor 1400W was determined for multiple intermediate end points characterizing BCG's direct effects on tumor cell biology. Activation of nuclear factor kappa B and nuclear factor (erythroid-derived 2)-like 2 signaling pathways, transactivation of genes, including p21, CD54, IL6, IL8, CXCL1, CXCL3, CCL20, and cytotoxicity, as measured by vital dye exclusion, lactate dehydrogenase release, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay were measured in response to BCG with and without iNOS inhibition.

RESULTS: Exposure of UC cells to BCG significantly increased both iNOS expression and NO production. Inhibition of iNOS activity with 1400W significantly inhibited BCG's direct biologic effect on UC cells for all of the end points evaluated.

CONCLUSIONS: iNOS expression, NO production, and the associated oxidative stress play a central role in the response of UC cells to BCG exposure. Manipulation of oxidative stress may afford an opportunity to enhance the antitumor effects of BCG.

Author List

Shah G, Zhang G, Chen F, Cao Y, Kalyanaraman B, See WA


Balaraman Kalyanaraman PhD Chair, Professor in the Biophysics department at Medical College of Wisconsin
William A. See MD Chair, Professor in the Urologic Surgery department at Medical College of Wisconsin


2-s2.0-84890805014   7 Citations

MESH terms used to index this publication - Major topics in bold

BCG Vaccine
Carcinoma, Transitional Cell
Cell Line, Tumor
Cell Proliferation
Chemokine CCL20
Chemokine CXCL1
Chemokines, CXC
Cyclin-Dependent Kinase Inhibitor p21
Gene Expression Regulation, Neoplastic
Intercellular Adhesion Molecule-1
NF-kappa B
Nitric Oxide
Nitric Oxide Synthase Type II
Reverse Transcriptase Polymerase Chain Reaction
Signal Transduction
Urinary Bladder Neoplasms
jenkins-FCD Prod-336 69ef4a6b262d135130251597d5d39873903802b5