Temporal proteome profiling of taxol-induced mitotic arrest and apoptosis.
Taxol (Paclitaxel) is a mitotic inhibitor widely used in cancer therapy. Temporal proteome profiling was performed to study changes of proteins during the different cellular states of HeLa cells caused by exposure to taxol. The changes of proteins over time could be associated with various cellular processes such as mitotic arrest, an intermediate between mitotic arrest and apoptosis, apoptosis, and late apoptosis. Calumenin, stress-induced phosphoprotein 1 (STIP1), and translationally controlled tumor protein (TCTP) were assigned to mitotic arrest and selected for further experiments using immunoblotting and subcellular fractionation. Calumenin translocated from membranes to the cytosol during mitotic arrest and late apoptosis, but was significantly reduced in the cytosol during apoptosis. Translocation of STIP1 to the nucleus was observed at apoptosis and to the cytoskeleton at late apoptosis. TCTP increased in the cytosol at mitotic arrest and in membranes at apoptosis. In addition, the quantitative time courses of Bim isoforms revealed differences between BimL and BimS in comparison with BimEL. In summary, temporal proteome profiling of HeLa cells incubated with taxol allowed the assignment of proteins to certain processes and additional experiments with complementary approaches enabled a more comprehensive understanding of spatial changes of selected proteins during mitotic arrest and apoptosis.
Bull
Vibeke H
VH
The Biotechnology Centre of Oslo, University of Oslo, Oslo, Norway.
Fargestad
Ellen M
EM
Strozynski
Margarita
M
Thiede
Bernd
B
D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D014408:Biomarkers, Tumor / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D006367:HeLa Cells; D006360:Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008938:Mitosis / Q000187:drug effects*; D017239:Paclitaxel / Q000494:pharmacology*; D025341:Principal Component Analysis; D020543:Proteome / Q000187:drug effects* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D050257:Tubulin Modulators / Q000494:pharmacology; D000090324:Tumor Protein, Translationally-Controlled 1
D000972:Antineoplastic Agents, Phytogenic; D014408:Biomarkers, Tumor; C510527:CALU protein, human; D002135:Calcium-Binding Proteins; D006360:Heat-Shock Proteins; D020543:Proteome; C079373:STIP1 protein, human; C000715647:TPT1 protein, human; D050257:Tubulin Modulators; D000090324:Tumor Protein, Translationally-Controlled 1; D017239:Paclitaxel