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Bitler BG , Fink LS , Wei Z , Peterson JR , Zhang R
A high-content screening assay for small-molecule modulators of oncogene-induced senescence
J Biomol Screen. 2013 Oct;18(9) :1054-61
PMID: 23733845    PMCID: PMC3870888     URL: https://www.ncbi.nlm.nih.gov/pubmed/23733845
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Abstract
Cellular senescence is a state of stable cell growth arrest. Activation of oncogenes such as RAS in mammalian cells typically triggers cellular senescence. Oncogene-induced senescence (OIS) is an important tumor suppression mechanism, and suppression of OIS contributes to cell transformation. Oncogenes trigger senescence through a multitude of incompletely understood downstream signaling events that frequently involve protein kinases. To identify target proteins required for RAS-induced senescence, we developed a small-molecule screen in primary human fibroblasts undergoing senescence induced by oncogenic RAS (H-Ras(G12V)). Using a high-content imaging system to monitor two hallmarks of senescence, senescence-associated beta-galactosidase activity expression and inhibition of proliferation, we screened a library of known small-molecule kinase inhibitors for those that suppressed OIS. Identified compounds were subsequently validated and confirmed using a third marker of senescence, senescence-associated heterochromatin foci. In summary, we have established a novel high-content screening platform that may be useful for elucidating signaling pathways mediating OIS by targeting critical pathway components.
Notes
Bitler, Benjamin G Fink, Lauren S Wei, Zhi Peterson, Jeffrey R Zhang, Rugang eng CA006927/CA/NCI NIH HHS/ R01GM083025/GM/NIGMS NIH HHS/ R01CA160331/CA/NCI NIH HHS/ CA010815/CA/NCI NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. J Biomol Screen. 2013 Oct;18(9):1054-61. doi: 10.1177/1087057113491827. Epub 2013 Jun 3.