Visualization of mitotic arrest of cell cycle with bioluminescence imaging in living animals

Mol Imaging Biol. 2013 Aug;15(4):431-40. doi: 10.1007/s11307-013-0619-x.

Abstract

Purpose: Visualization of the cell cycle in living subjects has long been a big challenge. The present study aimed to noninvasively visualize mitotic arrest of the cell cycle with an optical reporter in living subjects.

Procedures: An N-terminal cyclin B1-luciferase fusion construct (cyclin B-Luc) controlled by the cyclin B promoter, as a mitosis reporter, was generated. HeLa or HCT116 cells stably expressing cyclin B-Luc reporter were used to evaluate its cell cycle-dependent regulation and ubiquitination-mediated degradation. We also evaluated its feasibility to monitor the mitotic arrest caused by Taxotere both in vitro and in vivo.

Results: We showed that the cyclin B-Luc fusion protein was regulated in a cell cycle-dependent manner and accumulated in the mitotic phase (M phase) in cellular assays. The regulation of cyclin B-Luc reporter was mediated by proteasome ubiquitination. In the present study, in vitro imaging showed that antimitotic reagents like Taxotere upregulated the reporter through cell cycle arrest in the M phase. Noninvasive longitudinal bioluminescence imaging further demonstrated an upregulation of the reporter consistent with mitotic arrest induced in tumor xenograft models. Induction of this reporter was also observed with a kinesin spindle protein inhibitor, which causes cell cycle blockage in the M phase.

Conclusions: Our results demonstrate that the cyclin B-Luc reporter can be used to image whether compounds are capable, in vivo, of causing an M phase arrest and/or altering cyclin B turnover. This reporter can also be potentially used in high-throughput screening efforts aimed at discovering novel molecules that will cause cell cycle arrest at the M phase in cultivated cell lines and animal models.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology
  • Cell Cycle Checkpoints* / drug effects
  • Cyclin B1 / metabolism
  • Docetaxel
  • HCT116 Cells
  • HeLa Cells
  • Histones / metabolism
  • Humans
  • Luciferases / metabolism
  • Luminescence*
  • Mice
  • Mitosis* / drug effects
  • Molecular Imaging / methods*
  • Nocodazole / pharmacology
  • Phosphorylation / drug effects
  • Promoter Regions, Genetic / genetics
  • Protein Stability / drug effects
  • Proteolysis / drug effects
  • RNA, Small Interfering / metabolism
  • Recombinant Fusion Proteins / metabolism
  • Subcutaneous Tissue / drug effects
  • Taxoids / pharmacology
  • Ubiquitination / drug effects
  • Xenograft Model Antitumor Assays

Substances

  • Antineoplastic Agents
  • Cyclin B1
  • Histones
  • RNA, Small Interfering
  • Recombinant Fusion Proteins
  • Taxoids
  • Docetaxel
  • Luciferases
  • Nocodazole