Screening of cytotoxic or cytostatic flavonoids with quantitative Fluorescent Ubiquitination-based Cell Cycle Indicator-based cell cycle assay

• Published in: Royal Society open science Vol. 5; no. 12; p. 181303
• Main Authors: Go, Young-Hyun, Lee, Hyo-Ju, Kong, Hyeon-Joon, Jeong, Ho-Chang, Lee, Dong Young, Hong, Soon-Ki, Sung, Sang Hyun, Kwon, Ok-Seon, Cha, Hyuk-Jin
• Format: Journal Article
• Language: English
• Published: England The Royal Society 01.12.2018
• Subjects: cell cycle; Cellular and Molecular Biology; flavonoid; fucci; g2 arrest; sar; screening; FUCCI; screening; flavonoid; G2 arrest; SAR; cell cycle
• ISSN: 2054-5703; 2054-5703
• DOI: 10.1098/rsos.181303

The Fluorescent Ubiquitination-based Cell Cycle Indicator (FUCCI) system can be used not only to study gene expression at a specific cell cycle stage, but also to monitor cell cycle transitions in real time. In this study, we used a single clone of FUCCI-expressing HeLa cells (FUCCI-HeLa cells) and monitored the cell cycle in individual live cells over time by determining the ratios between red fluorescence (RF) of RFP-Cdt1 and green fluorescence (GF) of GFP-Geminin. Cytotoxic and cytostatic compounds, the latter of which induced G2 or mitotic arrest, were identified based on periodic cycling of the RF/GF and GF/RF ratios in FUCCI-HeLa cells treated with anti-cancer drugs. With this cell cycle monitoring system, ten flavonoids were screened. Of these, apigenin and luteolin, which have a flavone backbone, were cytotoxic, whereas kaempferol, which has a flavonol backbone, was cytostatic and induced G2 arrest. In summary, we developed a system to quantitatively monitor the cell cycle in real time. This system can be used to identify novel compounds that modulate the cell cycle and to investigate structure-activity relationships.