Cytotoxicity of withaferin A in glioblastomas involves induction of an oxidative stress-mediated heat shock response while altering Akt/mTOR and MAPK signaling pathways

Investigational New Drugs volume 31, pages545–557(2013)Cite this article

Summary

Withaferin A (WA), a steroidal lactone derived from the plant Vassobia breviflora, has been reported to have anti-proliferative, pro-apoptotic, and anti-angiogenic properties against cancer growth. In this study, we identified several key underlying mechanisms of anticancer action of WA in glioblastoma cells. WA was found to inhibit proliferation by inducing a dose-dependent G2/M cell cycle arrest and promoting cell death through both intrinsic and extrinsic apoptotic pathways. This was accompanied by an inhibitory shift in the Akt/mTOR signaling pathway which included diminished expression and/or phosphorylation of Akt, mTOR, p70 S6K, and p85 S6K with increased activation of AMPKα and the tumor suppressor tuberin/TSC2. Alterations in proteins of the MAPK pathway and cell surface receptors like EGFR, Her2/ErbB2, and c-Met were also observed. WA induced an N-acetyl-L-cysteine-repressible enhancement in cellular oxidative potential/stress with subsequent induction of a heat shock stress response primarily through HSP70, HSP32, and HSP27 upregulation and HSF1 downregulation. Taken together, we suggest that WA may represent a promising chemotherapeutic candidate in glioblastoma therapy warranting further translational evaluation.

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Acknowledgments

We would like to thank Dr. Jann Sarkaria of the Mayo Clinic (Rochester, MN) and Dr. John Ohlfest of the University of Minnesota (Minneapolis, MN) for generously providing the cell lines utilized. We would also like to thank the KUMC flow core facility for utilization of its resources as established by a generous endowment from the Hall Foundation and by NIH Grant Number P20 RR016443 from the COBRE program of the National Center for Research Resources

Conflict of interest

None declared

Role of funding sources

This work was made possible by grant support from the National Institutes of Health (NIH-COBRE P20 RR015563 P.I. B. Timmermann), the Institute for Advancing Medical Innovation (PI: MS Cohen), and a University of Kansas Cancer Center, Summer Student Training Program grant (PT Grogan).

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Affiliations

  1. Department of Pharmacology, Toxicology, and Therapeutics, University of Kansas Medical Center, Kansas, KS, 66160, USA

    Patrick T. Grogan & Mark S. Cohen

  2. Department of Surgery, University of Michigan Hospital and Health Systems, Ann Arbor, MI, 48109, USA

    Patrick T. Grogan & Mark S. Cohen

  3. Department of Surgery, University of Kansas Medical Center, Kansas, KS, 66160, USA

    Kristina D. Sleder & Abbas K. Samadi

  4. Department of Medicinal Chemistry, University of Kansas School of Pharmacy, Lawrence, KS, 66045, USA

    Huaping Zhang & Barbara N. Timmermann

  5. Section of General Surgery, University of Michigan Hospital and Health Systems, 2920K Taubman Center, SPC 5331, 1500 E. Medical Center Drive, Ann Arbor, MI, 48109-5331, USA

    Mark S. Cohen

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  1. Patrick T. Grogan
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  2. Kristina D. Sleder
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  3. Abbas K. Samadi
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  4. Huaping Zhang
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  5. Barbara N. Timmermann
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  6. Mark S. Cohen
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Correspondence to Mark S. Cohen.

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Grogan, P.T., Sleder, K.D., Samadi, A.K. et al. Cytotoxicity of withaferin A in glioblastomas involves induction of an oxidative stress-mediated heat shock response while altering Akt/mTOR and MAPK signaling pathways. Invest New Drugs 31, 545–557 (2013). https://doi.org/10.1007/s10637-012-9888-5

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Keywords

  • Withaferin A
  • Glioblastoma multiforme
  • Oxidative stress
  • Heat shock response
  • Akt/mTOR pathway
  • MAPK pathway