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J Biomol Screen. 2013 Oct;18(9):1062-71. doi: 10.1177/1087057113493804. Epub 2013 Jun 20.

Differential determinants of cancer cell insensitivity to antimitotic drugs discriminated by a one-step cell imaging assay.

Author information

1
1Department of Systems Biology, Harvard Medical School, Boston, MA, USA.

Abstract

Cancer cells can be drug resistant due to genetic variation at multiple steps in the drug response pathway, including drug efflux pumping, target mutation, and blunted apoptotic response. These are not discriminated by conventional cell survival assays. Here, we report a rapid and convenient high-content cell-imaging assay that measures multiple physiological changes in cells responding to antimitotic small-molecule drugs. Our one-step, no-wash assay uses three dyes to stain living cells and is much more accurate for scoring weakly adherent mitotic and apoptotic cells than conventional antibody-based assays. We profiled responses of 33 cell lines to 8 antimitotic drugs at multiple concentrations and time points using this assay and deposited our data and assay protocols into a public database (http://lincs.hms.harvard.edu/). Our data discriminated between alternative mechanisms that compromise drug sensitivity to paclitaxel and revealed an unexpected bell-shaped dose-response curve for BI2536, a highly selective inhibitor of Polo-like kinases. Our approach can be generalized, is scalable, and should therefore facilitate identification of molecular biomarkers for mechanisms of drug insensitivity in high-throughput screens and other assays.

KEYWORDS:

antimitotic drugs; cancer cells; drug sensitivity; high-content screening; image analysis; live cell imaging assay

PMID:
23788527
PMCID:
PMC3783590
DOI:
10.1177/1087057113493804
[Indexed for MEDLINE]
Free PMC Article
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