Variation in the kinetics of caspase-3 activation, Bcl-2 phosphorylation and apoptotic morphology in unselected human ovarian cancer cell lines as a response to docetaxel

• Published in: Biochemical pharmacology 2002-02, Vol.63 (4), p.733-743
• Main Authors: Kolfschoten, Geertruida M., Hulscher, Theresia M., Duyndam, Monique C.A., Pinedo, Herbert M., Boven, Epie
• Format: Article
• Language: English
• Subjects: Antineoplastic agents; Antineoplastic Agents, Phytogenic - pharmacology; Apoptosis; Bax; Bcl-2; Biological and medical sciences; Caspase 3; Caspases - metabolism; Cell Cycle - drug effects; Docetaxel; Enzyme Activation - drug effects; Female; General aspects; Human ovarian cancer cells; Humans; Kinetics; Medical sciences; Mitogen-Activated Protein Kinase 8; Mitogen-Activated Protein Kinase 9; Mitogen-Activated Protein Kinases - metabolism; Ovarian Neoplasms - pathology; Paclitaxel - analogs & derivatives; Paclitaxel - pharmacology; Pharmacology. Drug treatments; Phosphorylation; Proto-Oncogene Proteins c-bcl-2 - metabolism; Taxoids; Tumor Cells, Cultured
• ISSN: 0006-2952; 1873-2968
• DOI: 10.1016/S0006-2952(01)00895-4

Paclitaxel is able to cause cell death through the induction of apoptosis. Cell death characteristics for docetaxel have not yet been described in detail. We investigated four unselected human ovarian cancer cell lines for the sensitivity to a 1 hr exposure to docetaxel and calculated the concentrations inhibiting 50% ( ic 50) and 90% ( ic 90) of cell growth. Of the cell lines A2780, H134, IGROV-1 (all wild-type p53) and OVCAR-3 (mutant, mt p53) A2780 was most sensitive and OVCAR-3 least sensitive. Equitoxic drug concentrations representing ic 90 values (25–510 nM) were applied for 1 hr to measure cell cycle distribution, DNA degradation, and to count apoptotic cell bodies and cells with multifragmented nuclei at various time-points after drug exposure. H134, IGROV-1 and OVCAR-3 showed a continued mitotic block up to at least 72 hr and prolonged presence of cells with multifragmented nuclei. High percentages of apoptosis were calculated at 48 hr and at later time-points. In contrast, A2780 cells accumulated in the S-phase of the cell cycle and apoptosis was hardly present. The changes in the expression levels of p53, p21/WAF1, Bax and Bcl-2, were not predictive for docetaxel-induced apoptosis. Caspase-3 activation occurred only in cells with accumulation in the G2/M phase starting as early as 8 hr in OVCAR-3. Prolonged Bcl-2 phosphorylation was evident in OVCAR-3, visible at 24 hr in H134 and IGROV-1, while this phenomenon did not occur in A2780. The mitogen-activated protein kinase pathway (JNKs/SAPKs or c-Jun N-terminal kinases/stress-activated protein kinases, JNK1/2; extracellular response kinase, ERK1/2; p38) did not seem to be directly involved in Bcl-2 phosphorylation or apoptosis. We conclude that docetaxel is able to activate caspase-3, induce Bcl-2 phosphorylation and apoptosis in cells that show a prolonged G2/M arrest, but cells may also die by a caspase-3-independent cell death mechanism.