Visualization of radiation-induced cell cycle-associated events in tumor cells expressing the fusion protein of Azami Green and the destruction box of human Geminin

Biochem Biophys Res Commun. 2009 Nov 20;389(3):426-30. doi: 10.1016/j.bbrc.2009.08.160. Epub 2009 Sep 2.

Abstract

Ionizing radiation (IR) influences cell cycle-associated events in tumor cells. We expressed the fusion protein of Azami Green (AG) and the destruction box plus nuclear localization signal of human Geminin, an inhibitor of DNA replication licensing factor, in oral tumor cells. This approach allowed us to visualize G2 arrest in living cells following irradiation. The combination of time-lapse imaging analysis allowed us to observe the nuclear envelope break down (NEBD) at early M phase, and disappearance of fluorescence (DF) at the end of M phase. The duration from NEBD to DF was not much affected in irradiated cells; however, most of daughter cells harbored double-strand breaks. Complete DF was also observed in cells exhibiting abnormal mitosis or cytokinesis. We conclude that the fluorescent Geminin probe could function as a stable cell cycle indicator irrespective of genome integrity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle / radiation effects*
  • Cell Cycle Proteins / analysis
  • Cell Cycle Proteins / biosynthesis
  • Cell Cycle Proteins / metabolism*
  • Cell Line, Tumor
  • Cell Nucleus / metabolism*
  • Cytokinesis / radiation effects
  • DNA Breaks, Double-Stranded
  • G2 Phase / radiation effects
  • Geminin
  • Green Fluorescent Proteins / analysis
  • Green Fluorescent Proteins / biosynthesis
  • Green Fluorescent Proteins / metabolism*
  • Humans
  • Mitosis / radiation effects
  • Neoplasms / metabolism*
  • Nuclear Localization Signals / metabolism
  • Recombinant Fusion Proteins / analysis
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / metabolism*

Substances

  • Cell Cycle Proteins
  • GMNN protein, human
  • Geminin
  • Nuclear Localization Signals
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins