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A Dual-Color Fluorescence-Based Platform to Identify Selective Inhibitors of Akt Signaling

AuthorsRosado, Aránzazu ; Zanella, Fabian; García, Beatriz; Carnero, Amancio ; Link, Wolfgang
KeywordsBase sequence
Coculture techniques
DNA primers
Protein Kinase inhibitors
Proto-oncogene proteins c-akt
Issue Date19-Mar-2008
PublisherPublic Library of Science
CitationPLoS ONE 3(3): e1823 (2008)
AbstractBackground 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.
Description8 páginas, 5 figuras.
Publisher version (URL)http://dx.doi.org/10.1371/journal.pone.0001823
Appears in Collections:(IBIS) Artículos
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