Unusual expression of red fluorescence at M phase induced by anti-microtubule agents in HeLa cells expressing the fluorescent ubiquitination-based cell cycle indicator (Fucci).
Plinabulin (NPI-2358) is a novel microtubule-depolymerizing agent. In HeLa cells, plinabulin arrests the cell-cycle at M phase and subsequently induces mitotic catastrophe. To better understand the effects on this compound on the cell-cycle, we used the fluorescent ubiquitination-based cell cycle indicator (Fucci), which normally enables G1 and S/G2/M cells to emit red and green fluorescence, respectively. When HeLa-Fucci cells were treated with 50 nM plinabulin, cells began to fluoresce both green and red in an unusual pattern; most cells exhibited the new pattern after 24 h of treatment. X-irradiation efficiently induced G2 arrest in plinabulin-treated cells and significantly retarded the emergence of the unusual pattern, suggesting that entering M phase is essential for induction of the pattern. By simultaneously visualizing chromosomes with GFP-histone H2B, we established that the pattern emerges after nuclear envelope breakdown but before metaphase. Pedigree assay revealed a significant relationship between the unusual expression and mitotic catastrophe. Nocodazole, KPU-133 (a more potent derivative of plinabulin), and paclitaxel also exerted similar effects. From these data, we conclude that the unusual pattern may be associated with dysregulation of late M phase-specific E3 ligase activity and mitotic catastrophe following treatment with anti-microtubule agents.
Section of Oral Radiation Oncology, Department of Oral Health Sciences, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Tokyo 113-8549, Japan.
Asumi
A
Honda-Uezono
Atsushi
A
Kaida
Yasuyuki
Y
Michi
Kiyoshi
K
Harada
Yoshiki
Y
Hayashi
Yoshio
Y
Hayashi
Masahiko
M
Miura
D002455:Cell Division / Q000187:drug effects*; D054659:Diketopiperazines / Q000494:pharmacology*; D005453:Fluorescence; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis / Q000096:biosynthesis*; D008870:Microtubules / Q000187:drug effects*; D008938:Mitosis / Q000187:drug effects*; D015739:Nocodazole / Q000494:pharmacology; D050257:Tubulin Modulators / Q000494:pharmacology*; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism
D054659:Diketopiperazines; D008164:Luminescent Proteins; C514351:NPI 2358; D050257:Tubulin Modulators; D044767:Ubiquitin-Protein Ligases; D015739:Nocodazole