Cytosolic Ca2+ shifts as early markers of cytotoxicity

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2013
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Wyrsch, Philippe
Blenn, Christian
Pesch, Theresa
Althaus, Felix R.
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The determination of the cytotoxic potential of new and so far unknown compounds as well as their metabolites is fundamental in risk assessment. A variety of strategic endpoints have been defined to describe toxin-cell interactions, leading to prediction of cell fate. They involve measurement of metabolic endpoints, bio-energetic parameters or morphological cell modifications. Here, we evaluated alterations of the free cytosolic Ca2+ homeostasis using the Fluo-4 dye and compared results with the metabolic cell viability assay Alamar Blue. We investigated a panel of toxins (As2O3, gossypol, H2O2, staurosporine, and titanium(IV)-salane complexes) in four different mammalian cell lines covering three different species (human, mouse, and African green monkey). All tested compounds induced an increase in free cytosolic Ca2+ within the first 5 s after toxin application. Cytosolic Ca2+ shifts occurred independently of the chemical structure in all tested cell systems and were persistent up to 3 h. The linear increase of free cytosolic Ca2+ within the first 5 s of drug treatment correlates with the EC25 and EC75 values obtained in Alamar Blue assays one day after toxin exposure. Moreover, a rise of cytosolic Ca2+ was detectable independent of induced cell death mode as assessed by caspase and poly(ADP-ribose) polymerase (PARP) activity in HeLa versus MCF-7 cells at very low concentrations. In conclusion, a cytotoxicity assay based on Ca2+ shifts has a low limit of detection (LOD), is less time consuming (at least 24 times faster) compared to the cell viability assay Alamar Blue and is suitable for high-troughput-screening (HTS).

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ISO 690WYRSCH, Philippe, Christian BLENN, Theresa PESCH, Sascha BENEKE, Felix R. ALTHAUS, 2013. Cytosolic Ca2+ shifts as early markers of cytotoxicity. In: Cell Communication and Signaling. 2013, 11, 11. eISSN 1478-811X. Available under: doi: 10.1186/1478-811X-11-11
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@article{Wyrsch2013Cytos-30587,
  year={2013},
  doi={10.1186/1478-811X-11-11},
  title={Cytosolic Ca<sup>2+</sup> shifts as early markers of cytotoxicity},
  volume={11},
  journal={Cell Communication and Signaling},
  author={Wyrsch, Philippe and Blenn, Christian and Pesch, Theresa and Beneke, Sascha and Althaus, Felix R.},
  note={Article Number: 11}
}
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