Document Detail


The pyruvic acid analog 3-bromopyruvate interferes with the tetrazolium reagent MTS in the evaluation of cytotoxicity.
MedLine Citation:
PMID:  20085459     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
3-Bromopyruvate (3BrPA) is a pyruvate analog known for its alkylating property. Recently, several reports have documented the antiglycolytic and anticancer effects of 3BrPA and its potential for therapeutic applications. 3BrPA-mediated cytotoxicity has been evaluated in vitro by various methods including tetrazolium salt (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide)-based assays such as MTT, MTS, and so on. However, growing body of evidences has shown that tetrazolium reagent may interfere with the test compounds. In this study, we investigated whether the tetrazolium reagent interferes with the assessment of 3BrPA cytotoxicity. The results of the tetrazolium-based MTS assay were compared with 3 distinct cell viability detection methods, that is, Trypan Blue staining, ATP depletion, and Annexin V staining in 2 different cell lines, Vx-2 and HepG2. The MTS assay data showed false positive results by indicating increased cell viability at 1 mM and 2 mM 3BrPA whereas the other cell viability assays demonstrated that both Vx-2 and HepG2 cells are not viable at the same treatment conditions. In order to validate the direct interaction of 3BrPA with MTS reagent, we tested cell-free media incubated with different concentrations of 3BrPA. The results of cell-free media showed an increase in absorbance in a dose-dependent manner confirming the interaction of MTS with 3BrPA. Thus, our data clearly demonstrate that 3BrPA interferes with the accuracy of MTS-based cytotoxicity evaluation. Hence, we suggest that employing multiple methods of biochemical as well as morphological cytotoxicity assays is critical to evaluate 3BrPA-mediated cell death.
Authors:
Shanmugasundaram Ganapathy-Kanniappan; Jean-Francois H Geschwind; Rani Kunjithapatham; Manon Buijs; Labiq H Syed; Pramod P Rao; Shinichi Ota; Mustafa Vali
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Assay and drug development technologies     Volume:  8     ISSN:  1557-8127     ISO Abbreviation:  Assay Drug Dev Technol     Publication Date:  2010 Apr 
Date Detail:
Created Date:  2010-05-05     Completed Date:  2010-07-21     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101151468     Medline TA:  Assay Drug Dev Technol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  258-62     Citation Subset:  IM    
Affiliation:
Russell H. Morgan Department of Radiology and Radiological Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287, USA.
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MeSH Terms
Descriptor/Qualifier:
Adenosine Triphosphate / metabolism
Animals
Annexin A5 / metabolism
Biological Assay
Cell Line
Cell Line, Tumor
Cell Survival / drug effects*
Coloring Agents
Dose-Response Relationship, Drug
Drug Screening Assays, Antitumor
Humans
Indicators and Reagents
Pyruvates / chemistry*
Rabbits
Tetrazolium Salts / chemistry*
Thiazoles / chemistry*
Chemical
Reg. No./Substance:
0/Annexin A5; 0/Coloring Agents; 0/Indicators and Reagents; 0/Pyruvates; 0/Tetrazolium Salts; 0/Thiazoles; 1113-59-3/bromopyruvate; 298-93-1/thiazolyl blue; 56-65-5/Adenosine Triphosphate

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