Cell-based screen for altered nuclear phenotypes reveals senescence progression in polyploid cells after Aurora kinase B inhibition

Mol Biol Cell. 2015 Sep 1;26(17):2971-85. doi: 10.1091/mbc.E15-01-0003. Epub 2015 Jul 1.

Abstract

Cellular senescence is a widespread stress response and is widely considered to be an alternative cancer therapeutic goal. Unlike apoptosis, senescence is composed of a diverse set of subphenotypes, depending on which of its associated effector programs are engaged. Here we establish a simple and sensitive cell-based prosenescence screen with detailed validation assays. We characterize the screen using a focused tool compound kinase inhibitor library. We identify a series of compounds that induce different types of senescence, including a unique phenotype associated with irregularly shaped nuclei and the progressive accumulation of G1 tetraploidy in human diploid fibroblasts. Downstream analyses show that all of the compounds that induce tetraploid senescence inhibit Aurora kinase B (AURKB). AURKB is the catalytic component of the chromosome passenger complex, which is involved in correct chromosome alignment and segregation, the spindle assembly checkpoint, and cytokinesis. Although aberrant mitosis and senescence have been linked, a specific characterization of AURKB in the context of senescence is still required. This proof-of-principle study suggests that our protocol is capable of amplifying tetraploid senescence, which can be observed in only a small population of oncogenic RAS-induced senescence, and provides additional justification for AURKB as a cancer therapeutic target.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aurora Kinase B / antagonists & inhibitors*
  • Aurora Kinase B / genetics
  • Cell Division
  • Cell Line
  • Cell Nucleus / drug effects
  • Cell Nucleus / enzymology
  • Cell Nucleus / genetics
  • Cellular Senescence / drug effects
  • Cellular Senescence / genetics
  • Chromosome Segregation
  • Cytokinesis / genetics
  • HeLa Cells
  • High-Throughput Screening Assays / methods
  • Humans
  • Mitosis / drug effects
  • Mitosis / genetics
  • Phenotype
  • Polyploidy*
  • Protein Kinase Inhibitors / pharmacology*
  • Small Molecule Libraries / pharmacology

Substances

  • Protein Kinase Inhibitors
  • Small Molecule Libraries
  • Aurora Kinase B