Down-regulation of Notch1 by gamma-secretase inhibition contributes to cell growth inhibition and apoptosis in ovarian cancer cells A2780

Biochem Biophys Res Commun. 2010 Feb 26;393(1):144-9. doi: 10.1016/j.bbrc.2010.01.103. Epub 2010 Feb 1.

Abstract

The release of Notch intracellular domain (NICD) is mediated by gamma-secretase. gamma-Secretase inhibitors have been shown to be potent inhibitors of NICD. We hypothesized that Notch1 is acting as an oncogene in ovarian cancer and that inhibition of Notch1 would lead to inhibition of cell growth and apoptotic cell death in ovarian cancer cells. In this study, expressions of Notch1 and hes1 in four human ovarian cancer (A2780, SKOV3, HO-8910, and HO-8910PM), and one ovarian surface (IOSE 144) cell lines were detected by Western blot and quantitative real-time RT-PCR. The effects of gamma-secretase inhibition (N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester, DAPT) were measured by MTT assay, flow cytometry, ELISA and colony-forming assay. Our results showed that Notch1 and hes1 were found in all the four human ovarian cancer and IOSE 144 cell lines, and they were significantly higher in ovarian cancer cells A2780 compared to another four ovarian cells. Down-regulation of Notch1 expression by DAPT was able to substantially inhibit cell growth, induce G1 cell cycle arrest and induce cell apoptosis in A2780 in dose- and time-dependent manner. In addition, hes1 was found to be down-regulated in dose- and time-dependent manner by DAPT in A2780. These results demonstrate that treatment with DAPT leads to growth inhibition and apoptosis of A2780 cells in dose- and time-dependent manner. These findings also support the conclusion that blocking of the Notch1 activity by gamma-secretase inhibitors represents a potentially attractive strategy of targeted therapy for ovarian cancer.

MeSH terms

  • Amyloid Precursor Protein Secretases / antagonists & inhibitors*
  • Amyloid Precursor Protein Secretases / metabolism
  • Antineoplastic Agents / pharmacology*
  • Apoptosis*
  • Basic Helix-Loop-Helix Transcription Factors / antagonists & inhibitors
  • Basic Helix-Loop-Helix Transcription Factors / biosynthesis
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Dipeptides / pharmacology
  • Female
  • Homeodomain Proteins / antagonists & inhibitors
  • Homeodomain Proteins / biosynthesis
  • Humans
  • Ovarian Neoplasms / enzymology*
  • Protease Inhibitors / pharmacology*
  • Receptor, Notch1 / antagonists & inhibitors*
  • Receptor, Notch1 / biosynthesis
  • Transcription Factor HES-1

Substances

  • Antineoplastic Agents
  • Basic Helix-Loop-Helix Transcription Factors
  • Dipeptides
  • Homeodomain Proteins
  • N-(N-(3,5-difluorophenacetyl)alanyl)phenylglycine tert-butyl ester
  • NOTCH1 protein, human
  • Protease Inhibitors
  • Receptor, Notch1
  • Transcription Factor HES-1
  • HES1 protein, human
  • Amyloid Precursor Protein Secretases