Biochemical and Biophysical Research Communications, Vol.389, No.3, 426-430, 2009
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
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 We conclude that the fluorescent Geminin probe could exhibiting abnormal mitosis or cytokinesis function as a stable cell cycle indicator irrespective of genome integrity. (C) 2009 Elsevier Inc. All rights reserved.
Keywords:Ionizing radiation;DNA damage;Geminin;G2 arrest;Mitotic spindle checkpoint;Mitotic catastrophe