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PLoS One. 2008 Mar 19;3(3):e1823. doi: 10.1371/journal.pone.0001823.

A dual-color fluorescence-based platform to identify selective inhibitors of Akt signaling.

Author information

1
Experimental Therapeutics Program, Centro Nacional de Investigaciones Oncologicas, Madrid, Spain.

Abstract

BACKGROUND:

Inhibition of Akt signaling is considered one of the most promising therapeutic strategies for many cancers. However, rational target-orientated approaches to cell based drug screens for anti-cancer agents have historically been compromised by the notorious absence of suitable control cells.

METHODOLOGY/PRINCIPAL FINDINGS:

In order to address this fundamental problem, we have developed BaFiso, a live-cell screening platform to identify specific inhibitors of this pathway. BaFiso relies on the co-culture of isogenic cell lines that have been engineered to sustain interleukin-3 independent survival of the parental Ba/F3 cells, and that are individually tagged with different fluorescent proteins. Whilst in the first of these two lines cell survival in the absence of IL-3 is dependent on the expression of activated Akt, the cells expressing constitutively-activated Stat5 signaling display IL-3 independent growth and survival in an Akt-independent manner. Small molecules can then be screened in these lines to identify inhibitors that rescue IL-3 dependence.

CONCLUSIONS/SIGNIFICANCE:

BaFiso measures differential cell survival using multiparametric live cell imaging and permits selective inhibitors of Akt signaling to be identified. BaFiso is a platform technology suitable for the identification of small molecule inhibitors of IL-3 mediated survival signaling.

PMID:
18350159
PMCID:
PMC2267216
DOI:
10.1371/journal.pone.0001823
[Indexed for MEDLINE]
Free PMC Article

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