Identification of novel Notch target genes in T cell leukaemia

Mol Cancer. 2009 Jun 9;8:35. doi: 10.1186/1476-4598-8-35.

Abstract

Background: Dysregulated Notch signalling is believed to play an important role in the development and maintenance of T cell leukaemia. At a cellular level, Notch signalling promotes proliferation and inhibits apoptosis of T cell acute lymphoblastic leukaemia (T-ALL) cells. In this study we aimed to identify novel transcriptional targets of Notch signalling in the T-ALL cell line, Jurkat.

Results: RNA was prepared from Jurkat cells retrovirally transduced with an empty vector (GFP-alone) or vectors containing constitutively active forms of Notch (N1DeltaE or N3DeltaE), and used for Affymetrix microarray analysis. A subset of genes found to be regulated by Notch was chosen for real-time PCR validation and in some cases, validation at the protein level, using several Notch-transduced T-ALL and non-T-ALL leukaemic cell lines. As expected, several known transcriptional target of Notch, such as HES1 and Deltex, were found to be overexpressed in Notch-transduced cells, however, many novel transcriptional targets of Notch signalling were identified using this approach. These included the T cell costimulatory molecule CD28, the anti-apoptotic protein GIMAP5, and inhibitor of DNA binding 1 (1D1).

Conclusion: The identification of such downstream Notch target genes provides insights into the mechanisms of Notch function in T cell leukaemia, and may help identify novel therapeutic targets in this disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CD28 Antigens / genetics
  • CD28 Antigens / metabolism
  • Down-Regulation
  • Flow Cytometry
  • GTP Phosphohydrolases / genetics
  • GTP Phosphohydrolases / metabolism
  • Gene Expression Profiling*
  • Humans
  • Inhibitor of Differentiation Protein 1 / genetics
  • Inhibitor of Differentiation Protein 1 / metabolism
  • Jurkat Cells
  • Leukemia, T-Cell / genetics*
  • Leukemia, T-Cell / metabolism*
  • Oligonucleotide Array Sequence Analysis
  • Receptors, Notch / genetics*
  • Receptors, Notch / metabolism
  • Reproducibility of Results
  • Signal Transduction / genetics*
  • Up-Regulation
  • Vascular Endothelial Growth Factor A / genetics
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • CD28 Antigens
  • ID1 protein, human
  • Inhibitor of Differentiation Protein 1
  • Receptors, Notch
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • GTP Phosphohydrolases