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Akt promotes chemoresistance in human ovarian cancer cells by modulating cisplatin-induced, p53-dependent ubiquitination of FLICE-like inhibitory protein

Oncogene volume 29, pages 11–25 (2010)Cite this article

Abstract

Although Akt is a determinant of cisplatin (cis-diaminedichloroplatinum (CDDP)) resistance in ovarian cancer cells, which is related in part to its inhibitory action on p53 activation, precisely how Akt confers CDDP resistance is unclear. In this study, we show that CDDP induced p53-dependent Fas-associated death domain-like interleukin-1β-converting enzyme (FLICE)-like inhibitory protein (FLIP) degradation in chemosensitive ovarian cancer cells but not their resistant counterparts. CDDP induced FLIP–p53–Itch interaction, colocalization and FLIP ubiquitination in chemosensitive but not chemoresistant ovarian cancer cells. Moreover, although activated Akt inhibited CDDP-induced FLIP degradation and apoptosis in sensitive cells, these responses were facilitated by dominant-negative Akt expression in chemoresistant cells. Inhibition of Akt function also facilitated p53–FLIP interaction and FLIP ubiquitination, which were attenuated by p53 silencing. These results suggest that Akt confers resistance, in part, by modulating CDDP-induced, p53-dependent FLIP ubiquitination. Understanding the precise etiology of chemoresistance may improve treatment for ovarian cancer.

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Abbreviations

CDDP:

cis-diaminedichloroplatinum

DED:

death effector domain

DMEM:

Dulbecco's modified Eagle's medium

DMSO:

dimethyl sulfoxide

DN-Akt:

dominant-negative Akt

FADD:

Fas-associated death domain

FBS:

fetal bovine serum

FLICE:

Fas-associated death domain-like interleukin-1β-converting enzyme

FLIP:

FLICE-like inhibitory protein

FLIPL:

long isoform of FLIP

FLIPS:

short isoform of FLIP

GAPDH:

glyceraldehyde phosphate dehydrogenase

GFP:

green fluorescent protein

MDM2:

murine double minute-2

MOI:

multiple of infection

PI3K:

phosphatidylinositol-3-OH-kinase

PMSF:

phenylmethylsulfonyl fluoride

RPMI-1640:

Roswell Park Memorial Institute 1640

siRNA:

small interfering RNA

SDS–PAGE:

SDS–polyacrylamide gel electrophoresis

TP53:

tumor suppressor protein 53

XIAP:

X-linked inhibitor of apoptosis protein

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Acknowledgements

This work was supported by grants from the Cancer Research Society (# 11181 to BKT) and the Canadian Institutes of Health Research (#MOP-79360 to RB), the WCU (World Class University) program through the Korea Science and Engineering Foundation funded by the National Research Foundation of Korea (R31-10056) and a New Investigator Award to RB. MRA was the recipient of a Ministry of Health and Medical Education Scholarship, Iran. We thank Dr Qiao Li (University of Ottawa, Canada) and Dr Kenneth Walsh (St Elizabeth's Medical Centre, Boston, MA, USA) for providing HA-tagged ubiquitin and adenoviral HA-tagged DN-Akt, respectively. A2780s-AAkt2 and A2780s-PHM6 cells were generously provided by Dr Jin Cheng (H Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA).

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Authors and Affiliations

  1. Departments of Cellular & Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada

    M R Abedini, E J Muller, R Bergeron & B K Tsang

  2. Department of Physiology and Pharmacology, Birjand University of Medical Sciences, Birjand, Iran

    M R Abedini

  3. Neuroscience Program, Ottawa Hospital Research Institute, Ottawa, Ontario, Canada

    E J Muller & R Bergeron

  4. Department of Psychiatry, University of Ottawa, Ottawa, Ontario, Canada

    R Bergeron

  5. Department of Medicine, University of Ottawa, Ottawa, Ontario, Canada

    D A Gray

  6. Departments of Biochemistry, Microbiology & Immunology, University of Ottawa, Ottawa, Ontario, Canada

    D A Gray

  7. Cancer Therapeutics Program, Ottawa Hospital Research Institute, Ottawa, Ontario, Canada

    D A Gray

  8. Departments of Obstetrics & Gynaecology, University of Ottawa, Ottawa, Ontario, Canada

    B K Tsang

  9. Chronic Disease Program, Ottawa Hospital Research Institute, Ottawa, Ontario, Canada

    B K Tsang

  10. Department of Agricultural Biotechnology, World Class University Major in Biomodulation, College of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea

    B K Tsang

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Correspondence to B K Tsang.

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Abedini, M., Muller, E., Bergeron, R. et al. Akt promotes chemoresistance in human ovarian cancer cells by modulating cisplatin-induced, p53-dependent ubiquitination of FLICE-like inhibitory protein. Oncogene 29, 11–25 (2010). https://doi.org/10.1038/onc.2009.300

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