Implication of the Akt2/survivin pathway as a critical target in paclitaxel treatment in human ovarian cancer cells

Cancer Lett. 2009 Jan 18;273(2):257-65. doi: 10.1016/j.canlet.2008.08.027. Epub 2008 Oct 7.


Purpose: Although multiple mechanisms have been implicated in paclitaxel (PTX)-induced resistance in ovarian cancer, recent evidence has suggested that Akt2 has an important role in the protection of cells from paclitaxel-induced apoptosis. In the present study, we investigated the role of the Akt2/survivin pathway in paclitaxel-induced resistance by a modified method to generate an effective shRNA vector.

Methods: We applied RNAi-mediated silencing techniques to investigate the mechanism of the Akt2/survivin pathway on PTX-induced resistance in ovarian cancer cells (A2780 and SKOV3). The expression of Akt2 and survivin mRNA and related protein levels were evaluated with semiquantitative real-time RT-PCR and western blot analysis, respectively. Inhibition of cell proliferation was determined by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay, and the induction of apoptosis was examined through flow cytometry (FACS) and Hoechst staining.

Results: Akt2 down-regulation sensitized ovarian cancer cells to paclitaxel-induced apoptosis, and inhibited survivin expression. We further demonstrated that suppressing the inhibition of survivin expression can induce the drug-resistance to paclitaxel. We introduced a modified vector to generate shRNA to induce RNA interference, which contained three U6 promoters to express different shRNAs; it severely reduced Akt2 gene expression and showed good specificity.

Conclusion: Our findings will aid in understanding the molecular mechanism of paclitaxel-induced resistance in ovarian cancer and facilitate the development of novel anti-neoplastic strategies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis
  • Base Sequence
  • Cell Line, Tumor
  • Cell Proliferation
  • Cell Separation
  • Female
  • Flow Cytometry
  • Humans
  • Inhibitor of Apoptosis Proteins
  • Microtubule-Associated Proteins / metabolism*
  • Molecular Sequence Data
  • Neoplasm Proteins / metabolism*
  • Ovarian Neoplasms / metabolism*
  • Paclitaxel / pharmacology*
  • Proto-Oncogene Proteins c-akt / metabolism*
  • RNA Interference
  • Survivin
  • Tetrazolium Salts / pharmacology
  • Thiazoles / pharmacology


  • BIRC5 protein, human
  • Inhibitor of Apoptosis Proteins
  • Microtubule-Associated Proteins
  • Neoplasm Proteins
  • Survivin
  • Tetrazolium Salts
  • Thiazoles
  • AKT2 protein, human
  • Proto-Oncogene Proteins c-akt
  • thiazolyl blue
  • Paclitaxel