Live imaging of spindle pole disorganization in docetaxel-treated multicolor cells

Low-dose laulimalide represents a novel molecular probe for investigating microtubule organization

“…Previous time-lapse studies showed that metaphase human breast cancer cells treated with low doses of docetaxel resulted in a multipolar phenotype, with spindle poles gradually elongating parallel to the plate until eventual pole fragmentation. 19 As a control for the laulimalide experiment, synchronized cells were therefore treated with docetaxel for up to 2 h and analyzed with IIF. The dominant phenotype was confirmed to be multipolar with extensive centrosome stretching and fragmentation ( Fig.…”

“…19,33 The outcome of premature aster formation ranges from mild effects on the spindle as seen with epothilone A, 37 where the bipolar spindle is retained, to severe as seen with paclitaxel, 12 epothilone B 12,37 and docetaxel, 19 where multipolar spindles are formed with inhibition of pole separation at higher doses. When compared with these other MT-stabilizing agents, laulimalide favors an intermediate spindle phenotype, as long as the drug is present before nuclear envelope breakdown.…”

“…Cells were considered to have an abnormal morphology if they contained an aberrant chromosome pattern and/or more than two microtubule asters. From the abnormal cells, EC 50 values of 0.38 nM were obtained results are compared with the microtubule stabilizer docetaxel, a taxoid with well-characterized cellular effects, [18][19][20] in an attempt to uncouple the fundamental property of MT stabilization, which laulimalide and docetaxel hold in common, from the site of stabilization, where the two drugs differ. 1 In this paper we show that laulimalide represents a class of MT stabilizer that is distinct from the taxoids, inducing notable differences in cellular phenotype relative to docetaxel throughout the cell cycle, with the exception of prophase/prometaphase.…”

Visualization of Aberrant Perinuclear Microtubule Aster Organization by Microtubule-Destabilizing Agents

“…We have constructed a multicolor cell line in which the centrosomes and chromosomes could be simultaneously visualized, 7) and using this cell line, we recently characterized the in vivo effect of the microtubule-stabilizing agents taxanes and epothilones on centrosomes at low concentrations that do not cause mitotic arrest. 8,9) Since centrosomes are the dominant microtubule organizing centers (MTOCs) for spindle formation, it is thought that microtubule-targeting agents disturb the spindle architecture by inhibiting microtubule organization. In the present study, to characterize in detail the involvement of microtubule organization in spindle assembly disturbance, we constructed another multicolor MDA-MB-435 cell line in which the centrosomes, spindle microtubules, and chromosomes can be simultaneously visualized with EGFP-Aurora A, mCherry--tubulin, and ECFP-histone H3.…”

Synergistic Anticancer Effect of Peptide‐Docetaxel Nanoassembly Targeted to Tubulin: Toward Development of Dual Warhead Containing Nanomedicine

“…The bundle like microtubule generally formed because of taxol and is considered as a characteristic property to indicate the anticancer effect of cancer cell . It has been represented by live cell imaging that docetaxel treatment induced either aberrant multipolar spindles of which two centrosomes are mislocalized at one pole . Here, microtubules were stained green using primary anti‐α‐tubulin antibody and FITC labeled secondary antibody.…”