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Involvement of p21 in mitotic exit after paclitaxel treatment in MCF-7 breast adenocarcinoma cell line
N Barboule1, P Chadebech, V Baldin
1IPBS, CNRS 205, Toulouse, France.
Abstract:
It has been shown recently that expression of p21 is enhanced by paclitaxel. This cytotoxic compound induces mitotic spindle damage resulting in blockade of the mitotic cell cycle associated or not with apoptotic cell death. In the present study, we showed that, in MCF-7 cells, paclitaxel induced accumulation of p21 in cells with a G2/M DNA content, corresponding to cells either in abnormal mitosis or in an interphase-like state (decondensed chromatin) with multiple nuclei. In MCF-7 cells, the increase in p21 was subsequent to the mitotic arrest and was associated with the exit from abnormal mitosis leading to formation of cells with micronuclei. In this cell line, we noted a relationship between the elevation of p21 expression and the inhibition of p34cdc2 activity. High levels of p21 protein were also found to be associated with inactive p34cdc2/cyclin B protein complex after treatment with paclitaxel. Treatment with p21 antisense oligonucleotide partially blocked induction of p21 expression by paclitaxel and significantly reduced survival of MCF-7 cells exposed to this agent. In NIH-OVCAR-3 cells, which are deficient in basal and paclitaxel-induced p21 expression, paclitaxel led to a prolonged activation of p34cdc2 and a delayed mitotic exit associated with apoptotic cell death. These observations suggest that p21 is not required for the mitotic arrest in response to paclitaxel, but argue in favor of a role for this inhibitor in facilitating the exit from abnormal mitosis. This effectively enhances cell survival after paclitaxel-induced spindle damage.
Insights
Paclitaxel enhances p21 expression, aiding exit from abnormal mitosis and improving cell survival after spindle damage. This inhibitor is crucial for cell fate decisions following mitotic arrest.
Area of Science:
- Molecular Biology
- Cell Biology
- Cancer Research
Background:
- Paclitaxel is a cytotoxic agent that damages mitotic spindles, causing cell cycle arrest and potentially apoptosis.
- Previous studies indicate paclitaxel enhances the expression of p21, a cell cycle inhibitor.
Purpose of the Study:
- To investigate the role of p21 in MCF-7 and NIH-OVCAR-3 cells following paclitaxel treatment.
- To elucidate the relationship between p21 expression, mitotic arrest, and cell survival after paclitaxel-induced spindle damage.
Main Methods:
- Treatment of MCF-7 and NIH-OVCAR-3 cells with paclitaxel.
- Analysis of p21 accumulation, p34cdc2 activity, and p34cdc2/cyclin B complex formation.
- Use of p21 antisense oligonucleotide to assess its impact on cell survival.
Main Results:
- In MCF-7 cells, paclitaxel induced p21 accumulation subsequent to mitotic arrest, correlating with abnormal mitosis exit and micronuclei formation.
- Elevated p21 expression was linked to inhibited p34cdc2 activity and inactive p34cdc2/cyclin B complex.
- In p21-deficient NIH-OVCAR-3 cells, paclitaxel caused prolonged p34cdc2 activation and delayed mitotic exit, leading to apoptosis.
Conclusions:
- p21 is not essential for paclitaxel-induced mitotic arrest but plays a role in facilitating exit from abnormal mitosis.
- p21 accumulation enhances cell survival after paclitaxel-induced spindle damage by promoting a proper exit from aberrant mitotic states.
- These findings highlight p21's critical function in determining cell fate following cytotoxic chemotherapy.