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

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.

• Authors: Mahito Sadaie, Christian Dillon, Masako Narita, Masashi Narita, Andrew R J Young, Claire J Cairney, Lauren S Godwin, Christopher J Torrance, Dorothy C Bennett, W Nicol Keith, Masashi Narita
• Journal: Molecular biology of the cell (Mol Biol Cell) Vol. 26 Issue 17 Pg. 2971-85 (Sep 01 2015) ISSN: 1939-4586 [Electronic] United States
• PMID: 26133385 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2015 Sadaie et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

Chemical References

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

Topics

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