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 exhibiting abnormal mitosis or cytokinesis. We conclude that the fluorescent Geminin probe could function as a stable cell cycle indicator irrespective of genome integrity.
Ishikawa
Mayuko
M
Oral Radiation Oncology, Department of Oral Restitution, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Ogihara
Yusuke
Y
Miura
Masahiko
M
D002453:Cell Cycle / Q000528:radiation effects*; D018797:Cell Cycle Proteins / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism*; D048749:Cytokinesis / Q000528:radiation effects; D053903:DNA Breaks, Double-Stranded; D016195:G2 Phase / Q000528:radiation effects; D064248:Geminin; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*; D006801:Humans; D008938:Mitosis / Q000528:radiation effects; D009369:Neoplasms / Q000378:metabolism*; D019913:Nuclear Localization Signals / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*
D018797:Cell Cycle Proteins; C577995:GMNN protein, human; D064248:Geminin; D019913:Nuclear Localization Signals; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins