Secondary stimulation from Bacillus Calmette-Guérin induced macrophages induce nitric oxide independent cell-death in bladder cancer cells

Tomas Thiel; Charlotta Ryk; Vicky Chatzakos; Katarina Hallén Grufman; Nasrin Bavand-Chobot; Jenny Flygare; N Peter Wiklund; Petra J de Verdier

doi:10.1016/j.canlet.2014.03.014

Secondary stimulation from Bacillus Calmette-Guérin induced macrophages induce nitric oxide independent cell-death in bladder cancer cells

Cancer Lett. 2014 Jun 28;348(1-2):119-25. doi: 10.1016/j.canlet.2014.03.014. Epub 2014 Mar 18.

Authors

Tomas Thiel¹, Charlotta Ryk¹, Vicky Chatzakos², Katarina Hallén Grufman¹, Nasrin Bavand-Chobot¹, Jenny Flygare³, N Peter Wiklund¹, Petra J de Verdier⁴

Affiliations

¹ Department of Urology, Karolinska University Hospital, Stockholm, Sweden; Urology Laboratory, Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden.
² Department of Urology, Karolinska University Hospital, Stockholm, Sweden; Urology Laboratory, Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden; Laboratory Medicine, Division of Clinical Chemistry, Karolinska Institutet and Karolinska University Hospital Huddinge, Stockholm, Sweden; Stockholm University, Department of Genetics, Microbiology and Toxicology, Stockholm, Sweden.
³ Laboratory Medicine, Division of Clinical Chemistry, Karolinska Institutet and Karolinska University Hospital Huddinge, Stockholm, Sweden.
⁴ Department of Urology, Karolinska University Hospital, Stockholm, Sweden; Urology Laboratory, Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden. Electronic address: petra.deverdier@ki.se.

PMID: 24657658
DOI: 10.1016/j.canlet.2014.03.014

Abstract

The anti-tumour mechanisms following Bacillus Calmette-Guérin (BCG) treatment of bladder-cancer remain largely unknown. Previous studies have shown involvement of nitric-oxide (NO) formation in the BCG-mediated effect. We analyzed the effects of macrophage secreted factors (MSFs) from BCG-stimulated RAW264.7 cells on the bladder-cancer cell line MBT2. Direct treatment with BCG did not induce NO in MBT2-cells whereas supernatant from BCG-stimulated macrophages increased NOS2 mRNA and protein expression, NO concentrations and cell-death. Blocking NO-synthesis with the NOS-inhibitor L-NAME did not affect levels of cell-death suggesting cytotoxic pathways involving other signalling molecules than NO. Several such candidate genes were identified in a microarray.

Keywords: Bacillus Calmette-Guérin; Cytokines; Nitric oxide; Nitric oxide synthase; Urinary bladder cancer.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antineoplastic Agents / therapeutic use*
BCG Vaccine / therapeutic use*
Cell Death / drug effects
Cell Line, Tumor
Culture Media, Conditioned / metabolism
Dose-Response Relationship, Drug
Enzyme Inhibitors / pharmacology
Gene Expression Profiling / methods
Humans
Macrophages / drug effects*
Macrophages / immunology
Macrophages / metabolism
Mice
Nitric Oxide / metabolism*
Nitric Oxide Synthase Type II / antagonists & inhibitors
Nitric Oxide Synthase Type II / genetics
Nitric Oxide Synthase Type II / metabolism
Oligonucleotide Array Sequence Analysis
Paracrine Communication / drug effects*
RNA, Messenger / metabolism
Signal Transduction / drug effects
Treatment Outcome
Up-Regulation
Urinary Bladder Neoplasms / drug therapy*
Urinary Bladder Neoplasms / genetics
Urinary Bladder Neoplasms / immunology
Urinary Bladder Neoplasms / metabolism
Urinary Bladder Neoplasms / pathology

Substances

Antineoplastic Agents
BCG Vaccine
Culture Media, Conditioned
Enzyme Inhibitors
RNA, Messenger
Nitric Oxide
NOS2 protein, human
Nitric Oxide Synthase Type II
Nos2 protein, mouse