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Comparison of three cytotoxicity tests in the evaluation of the cytotoxicity of a spermine analogue on human breast cancer cell lines

Holst, Martina LU and Oredsson, Stina LU (2005) In Toxicology in Vitro 19(3). p.379-387
Abstract
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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Apoptosis, Cell proliferation, Human breast cancer, Mitochondrial activity, Spermine analogue
in
Toxicology in Vitro
volume
19
issue
3
pages
379 - 387
publisher
Elsevier
external identifiers
  • pmid:15713545
  • wos:000227760800006
  • scopus:13844298327
ISSN
1879-3177
DOI
10.1016/j.tiv.2004.10.005
language
English
LU publication?
yes
id
d1b55631-e5dd-430c-a0d1-f513e5c28849 (old id 168004)
date added to LUP
2007-07-23 11:22:43
date last changed
2017-01-01 07:12:49
@article{d1b55631-e5dd-430c-a0d1-f513e5c28849,
  abstract     = {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<br/><br>
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<br/><br>
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.},
  author       = {Holst, Martina and Oredsson, Stina},
  issn         = {1879-3177},
  keyword      = {Apoptosis,Cell proliferation,Human breast cancer,Mitochondrial activity,Spermine analogue},
  language     = {eng},
  number       = {3},
  pages        = {379--387},
  publisher    = {Elsevier},
  series       = {Toxicology in Vitro},
  title        = {Comparison of three cytotoxicity tests in the evaluation of the cytotoxicity of a spermine analogue on human breast cancer cell lines},
  url          = {http://dx.doi.org/10.1016/j.tiv.2004.10.005},
  volume       = {19},
  year         = {2005},
}