Aurora kinase A inhibition leads to p73-dependent apoptosis in p53-deficient cancer cells

Altaf A. Dar, Abbes Belkhiri, Jeffrey Ecsedy, Alexander Zaika, Wael El-Rifai

Research output: Contribution to journalArticle

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Abstract

We investigated the role of Aurora kinase A (AURKA) in regulating p73-dependent apoptosis using the p53-deficient cancer cell lines H1299, TE7, and HCT116p53-/-. Overexpression of AURKA led to down-regulation of the TAp73-induced activation of the p53/p73-dependent luciferase reporter plasmid (pG13-luc). The reduction in the TAp73 transcription activity was confirmed by measuring the activity of luciferase reporters for p21/WAF1, and PUMA. The siRNA knockdown of endogenous AURKA reversed these effects and Western blot analysis showed a significant increase in the protein level of TAp73 and its downstream transcription targets, PUMA, NOXA, and p21/WAF1. The coexpression of AURKA together with TAp73 inhibited the activation of the pG13-luc, PUMA-luc, and p21/WAF1-luc reporter plasmids with reduction in the protein levels of TAp73 transcription targets. Treatment with AURKA-selective small molecule inhibitor MLN8054 led to a significant increase in the activities of pG13-luc, PUMA-luc, and p21/WAF1-luc reporter plasmids. This effect was accompanied by a significant increase in the mRNA and protein levels of several TAp73 transcription targets: p21/WAF1, PUMA, and NOXA. Flow cytometry cell cycle analysis, after MLN8054 treatment, showed more than a 2-fold increase in cell death. The apoptotic outcome was corroborated by showing an increase in cleaved caspase-3 protein levels by Western blot. Using terminal deoxynucleotidyl- transferase-mediated dUTP nick-end labeling assay, we showed that the expression of dominant-negative mutant TAp73 expression plasmid (p73DD) counter-acted the MLN8054-induced cell death. Taken together, our results indicate that AURKA regulates TAp73-dependent apoptosis and highlight the potential of the AURKA inhibitor MLN8054 in treating cancers that are defective in p53 signaling.

Original languageEnglish (US)
Pages (from-to)8998-9004
Number of pages7
JournalCancer Research
Volume68
Issue number21
DOIs
StatePublished - Nov 1 2008
Externally publishedYes

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Aurora Kinase A
Apoptosis
Plasmids
Neoplasms
Luciferases
Proteins
Cell Death
Western Blotting
DNA Nucleotidylexotransferase
Caspase 3
Small Interfering RNA
Cell Cycle
Flow Cytometry
Down-Regulation
Cell Line
Messenger RNA
MLN8054

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Aurora kinase A inhibition leads to p73-dependent apoptosis in p53-deficient cancer cells. / Dar, Altaf A.; Belkhiri, Abbes; Ecsedy, Jeffrey; Zaika, Alexander; El-Rifai, Wael.

In: Cancer Research, Vol. 68, No. 21, 01.11.2008, p. 8998-9004.

Research output: Contribution to journalArticle

Dar, Altaf A. ; Belkhiri, Abbes ; Ecsedy, Jeffrey ; Zaika, Alexander ; El-Rifai, Wael. / Aurora kinase A inhibition leads to p73-dependent apoptosis in p53-deficient cancer cells. In: Cancer Research. 2008 ; Vol. 68, No. 21. pp. 8998-9004.
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