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Protein kinase Cɛ confers resistance of MCF-7 cells to TRAIL by Akt-dependent activation of Hdm2 and downregulation of p53

Oncogene volume 27, pages 3957–3966 (2008)Cite this article

Abstract

Protein kinase Cɛ (PKCɛ) acts as an antiapoptotic protein and inhibits tumor necrosis factor-α (TNF)-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in MCF-7 breast cancer cells. Members of the TNF receptor superfamily trigger apoptosis independent of the tumor suppressor protein p53, which primarily affects DNA damage-induced apoptosis. We have previously shown that PKCɛ acts upstream of Akt to inhibit receptor-initiated cell death. Since Akt can regulate p53, we have examined the involvement of p53 in PKCɛ-mediated TRAIL resistance. Overexpression of PKCɛ in MCF-7 cells (MCF-7/PKCɛ) caused a decrease in p53 and an increase in human homolog of murine double minute 2 (Hdm2) and phospho-Hdm2. Depletion of p53 by siRNA attenuated, whereas depletion of Hdm2 enhanced TRAIL-mediated apoptosis. Knockdown of Akt decreased Hdm2 phosphorylation, increased p53 level and potentiated TRAIL-induced cell death. Depletion of PKCɛ from MCF-7 cells caused an increase in p53, whereas knockdown of p53 caused a decrease in Bid mRNA. Depletion of Akt from MCF-7/PKCɛ cells resulted in an increase in p53 and Bid. These results suggest that PKCɛ mediates TRAIL resistance by Akt-mediated phosphorylation of Hdm2 resulting in suppression of p53 expression and downregulation of Bid in MCF-7 breast cancer cells.

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Abbreviations

Hdm2:

human homolog of Mdm2

Mdm2:

murine double minute 2

PARP:

poly (ADP-ribose) polymerase

PKB:

protein kinase B

PKC:

protein kinase C

SDS–PAGE:

sodium dodecyl sulfate–polyacrylamide gel electrophoresis

TNF:

tumor necrosis factor-α

TRAIL:

TNF-related apoptosis-inducing ligand

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Acknowledgements

We thank Chandreyi Basu for technical assistance and Dr Arvind Virmani and Dr Adi Gazdar for providing us HCC1806 cells. This work is supported by the grant CA71727 from the NCI.

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Author notes

  1. E Shankar and U Sivaprasad: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Biology and Immunology, University of North Texas Health Science Center, Fort Worth, TX, USA

    E Shankar, U Sivaprasad & A Basu

  2. Institute for Cancer Research, University of North Texas Health Science Center, Fort Worth, TX, USA

    E Shankar, U Sivaprasad & A Basu

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  1. E Shankar
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Correspondence to A Basu.

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Shankar, E., Sivaprasad, U. & Basu, A. Protein kinase Cɛ confers resistance of MCF-7 cells to TRAIL by Akt-dependent activation of Hdm2 and downregulation of p53. Oncogene 27, 3957–3966 (2008). https://doi.org/10.1038/onc.2008.39

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