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Paclitaxel induces inactivation of p70 S6 kinase and phosphorylation of Thr421 and Ser424 via multiple signaling pathways in mitosis

Oncogene volume 22, pages 484–497 (2003)Cite this article

Abstract

The 70 kDa ribosomal S6 kinase (p70S6K) is important for cell growth and survival. Activation of p70S6K requires sequential phosphorylation of multiple serine and threonine sites often triggered by growth factors and hormones. Here, we report that paclitaxel, a microtubule-damaging agent, induces phosphorylation of p70S6K at threonine 421 and serine 424 (T421/S424) in a concentration- and time-dependent manner in multiple breast and ovarian cancer cell lines demonstrated by a T421/S424 phospho-p70S6K antibody. Phosphoamino-acid analysis and Western blot analysis by serine-/threonine-specific antibodies further confirms that both serine and threonine residues are phosphorylated in p70S6K following treatment with paclitaxel. Paclitaxel-induced p70S6KT421/S424 phosphorylation requires both de novo RNA and protein synthesis via multiple signaling pathways including ERK1/2 MAP kinase, JNK, PKC, Ca++, PI3K, and mammalian target of rapamycin (mTOR). Despite phosphorylation of p70S6KT421/S424, paclitaxel inactivates this kinase in a concentration- and time-dependent manner as illustrated by in vitro kinase assay. Inhibitors of mTOR, PI3K, and Ca++ impair p70S6K activity, whereas inhibitors of JNK and PKC stimulate p70S6K activity. Inhibition of PKC and JNK prevents paclitaxel-induced p70S6K inactivation. Moreover, the paclitaxel-induced phosphorylation and low activity of p70S6K mainly occurs during mitosis. In summary, paclitaxel is able to induce p70S6KT421/S424 phosphorylation and decrease its activity in mitotic cells via multiple signaling pathways. Our data suggest that paclitaxel-induced p70S6KT421/S424 phosphorylation and kinase inactivation are differentially regulated. Our data also indicate that paclitaxel may exert its antitumor effect, at least in part, via inhibition of p70S6K.

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Acknowledgements

We sincerely thank Drs J Avruch and DJ Templeton for providing mutant plasmids of p70S6K and Dr Z Siddik for providing 2008 cells. We also thank Karen Ramirez at Flow Cytometry Core Laboratory (Smith Research Building) of MD Anderson Cancer Center for her expert assistance with flow cytometry analysis.

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Authors and Affiliations

  1. Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, 77030, TX, USA

    Xiao-Feng Le, Walter N Hittelman, Amanda McWatters & Robert C Bast Jr

  2. Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, 77030, TX, USA

    Jiaxin Liu

  3. Diagnostic Radiology, The University of Texas MD Anderson Cancer Center, Houston, 77030, TX, USA

    Chun Li

  4. Molecular Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, 77030, TX, USA

    Gordon B Mills

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  1. Xiao-Feng Le
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Correspondence to Robert C Bast Jr.

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Our research was supported in part by a Grant CA-39930 (to RCB) from the National Institutes of Health and by an Institutional Research Grant IRG-3721206 (to XFL) from the University of Texas MD Anderson Cancer Center

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Le, XF., Hittelman, W., Liu, J. et al. Paclitaxel induces inactivation of p70 S6 kinase and phosphorylation of Thr421 and Ser424 via multiple signaling pathways in mitosis. Oncogene 22, 484–497 (2003). https://doi.org/10.1038/sj.onc.1206175

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