A high-content screening assay for small-molecule modulators of oncogene-induced senescence

Benjamin G. Bitler, Lauren S. Fink, Zhi Wei, Jeffrey R. Peterson, Rugang Zhang

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

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-RasG12V). Using a high-content imaging system to monitor two hallmarks of senescence, senescence-associated β-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.

Original languageEnglish (US)
Pages (from-to)1054-1061
Number of pages8
JournalJournal of Biomolecular Screening
Volume18
Issue number9
DOIs
StatePublished - Oct 2013

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Molecular Medicine
  • Pharmacology
  • Drug Discovery

Keywords

  • cancer and cancer drugs
  • cell-based assays
  • high-content screening
  • kinases

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