Comparison of three cytotoxicity tests in the evaluation of the cytotoxicity of a spermine analogue on human breast cancer cell lines
Author
Summary, in English
Using three cytotoxicity assays, we have investigated the effect of the spermine analogue N1,N11-diethylnorspermine (DENSPM)on four human breast cancer cell lines with different known genetic lesions. Cells were seeded in 96 well plates and DENSPM was added 24 h later to give final concentrations from 0.1 to 100 microM. At 24, 48 and 72 h of treatment, the protein content was determined with a modified Lowry assay. Mitochondrial activity was determined with the AlamarBlue and MTT assays. These two
assays differ with respect to where in the electron transport chain the reduction of the substrate takes place. Treatment with increasing concentrations of DENSPM resulted in differential responses in the four cell lines. There was a good of agreement between the protein content and the MTT assay showing increased negative effect with increased dose of DENSPM. The AlamarBlue assay on
the other hand showed a stimulation of substrate reduction compared to control at DENSPM concentrations that were inhibitory according to the protein content and MTT assay. Thus, the data clearly show that the MTT and AlamarBlue assays are not equivalent. Importantly, the AlamarBlue assay presumably also reflects cytoplasmic reduction of the substrate through DENSPM-induced mechanisms.
assays differ with respect to where in the electron transport chain the reduction of the substrate takes place. Treatment with increasing concentrations of DENSPM resulted in differential responses in the four cell lines. There was a good of agreement between the protein content and the MTT assay showing increased negative effect with increased dose of DENSPM. The AlamarBlue assay on
the other hand showed a stimulation of substrate reduction compared to control at DENSPM concentrations that were inhibitory according to the protein content and MTT assay. Thus, the data clearly show that the MTT and AlamarBlue assays are not equivalent. Importantly, the AlamarBlue assay presumably also reflects cytoplasmic reduction of the substrate through DENSPM-induced mechanisms.
Department/s
Publishing year
2005
Language
English
Pages
379-387
Publication/Series
Toxicology in Vitro
Volume
19
Issue
3
Document type
Journal article
Publisher
Elsevier
Topic
- Zoology
Keywords
- Apoptosis
- Cell proliferation
- Human breast cancer
- Mitochondrial activity
- Spermine analogue
Status
Published
ISBN/ISSN/Other
- ISSN: 1879-3177