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Fluorescence kinetics in HeLa cells after treatment with cell cycle arrest inducers visualized with Fucci (fluorescent ubiquitination‐based cell cycle indicator)
Author(s) -
Kaida Atsushi,
Sawai Naoki,
Sakaguchi Kengo,
Miura Masahiko
Publication year - 2011
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1042/cbi20100643
Subject(s) - nocodazole , hela , cell cycle , fluorescence , microbiology and biotechnology , flow cytometry , kinetics , cell , biophysics , green fluorescent protein , cell cycle checkpoint , mitosis , biology , chemistry , biochemistry , gene , physics , quantum mechanics , cytoskeleton
Fucci (fluorescent ubiquitination‐based cell cycle indicator) is able to visualize dynamics of cell cycle progression in live cells; G 1 ‐ and S‐/G 2 ‐/M‐phase cells expressing Fucci emit red and green fluorescence, respectively. This system could be applied to cell kinetic analysis of tumour cells in the field of cancer therapy; however, it is still unclear how fluorescence kinetics change after various treatments, including exposure to anticancer agents. To explore this, we arrested live HeLa cells expressing the Fucci probes at various cell cycle stages and observed the fluorescence, in conjunction with flow cytometric analysis. X‐irradiation, HU (hydroxyurea) and nocodazole arrest cells at G 2 /M boundary, early S‐phase and early M‐phase, respectively. Although X‐irradiation and HU treatment induced similar accumulation kinetics of green fluorescent cells, nocodazole treatment induced an abnormal red fluorescence at M phase, followed by accumulation of both red and green fluorescent cells with 4N DNA content. We conclude that certain agents that disrupt normal cell cycle regulation could cause unexpected fluorescence kinetics in the Fucci system.