The colchicine derivative CT20126 shows a novel microtubule-modulating activity with apoptosis

Abstract

New colchicine analogs have been synthesized with the aim of developing stronger potential anticancer activities. Among the analogs, CT20126 has been previously reported to show immunosuppressive activities. Here, we report that CT20126 also shows potential anticancer effects via an unusual mechanism: the modulation of microtubule integrity and cell cycle arrest at the G2/M phase before apoptosis. When we treated COS-7 cells with CT20126 (5 lM), the normal thread-like microtubules were disrupted into tubulin dimers within 10min and thereafter repolymerized into short, thick filaments. In contrast, cells treated with the same concentration of colchicine exhibited microtubule depolymerization after 20min and never underwent repolymerization. Furthermore, optical density (OD) analysis (350nm) with purified tubulin showed that CT20126 had a higher repolymerizing activity than that of Taxol, a potent microtubule-polymerizing agent. These results suggest that the effects of CT20126 on microtubule integrity differ from those of colchicine: the analog first destabilizes microtubules and then stabilizes the disrupted tubulins into short, thick polymers. Furthermore, CT20126 induced a greater level of apoptotic activity in Jurkat T cells than colchicine (assessed by G2/M arrest, caspase-3 activation and cell sorting). At 20nM, CT20126 induced 47% apoptosis among Jurkat T cells, whereas colchicine induced only 33% apoptosis. Our results suggest that the colchicine analog CT20126 can potently induce apoptosis by disrupting microtubule integrity in a manner that differs from that of colchicine or Taxol. Experimental & Molecular Medicine (2013) 45, e19; doi:10.1038/emm.2013.38; published online 19 April 2013 Keywords: apoptosis; cell cycle arrest; colchicine analog; CT20126; microtubule disruption; tubulin acetylation INTRODUCTION Microtubules, which are structural components of the cytoskeleton, are composed of a- and b-tubulin heterodimers and play key roles in many biological events, including the development and maintenance of cell shape, intracellular transport, cell division and cell growth.1,2 A subset of microtubules called spindle fibers support proper cell division by pulling the chromosomes to either side of the dividing cell. In