Level of Notch activation determines the effect on growth and stem cell-like features in glioblastoma multiforme neurosphere cultures

Abstract

Background: Brain cancer stem-like cells (bCSC) are cancer cells with neural stem cell (NSC)-like properties found in glioblastoma multiforme (GBM) and they are assigned a central role in tumor initiation, progression and relapse. The Notch pathway is important for maintenance and cell fate decisions in the normal NSC population. Notch signaling is often deregulated in GBM and recent results suggest that this pathway plays a significant role in bCSC as well. We therefore wished to further elucidate the role of Notch activation in GBM-derived bCSC.

 

Methods: Human-derived GBM xenograft cells were cultured as NSC-like neurosphere cultures. Notch modulation was accomplished either by blocking the pathway using the γ-secretase inhibitor DAPT or by activating it by transfecting the cells with the constitutive active Notch-1 receptor.

Results: GBM neurosphere cultures with high endogenous Notch activation displayed sensitivity toward Notch inhibition with regard to tumorigenic features as demonstrated by increased G₀/G₁ population and reduced colony formation capacity. Of the NSC-like characteristics, only the primary sphere forming potential was affected, while no effect was observed on self-renewal or differentiation. In contrast, when Notch signaling was activated a decrease in the G₀/G₁ population and an enhanced capability of colony formation was observed, along with increased self-renewal and de-differentiation.

Conclusion: Based on the presented results we propose that active Notch signaling plays a role for cell growth and stem cell-like features in GBM neurosphere cultures and that Notch-targeted anti-bCSC treatment could be feasible for GBM patients with high endogenous Notch pathway activation.

Keywords: :

• glioblastoma multiforme
• neurosphere cultures
• brain cancer stem-like cells
• Notch signaling
• Notch activity
• DAPT
• ICN-1

Disclosure of Potential Conflicts of Interest

No potential conflict of interest was disclosed.

Acknowledgments

We thank statistician Susanne Rosthøj for assistance performing the statistical analysis and technician Pia Pedersen for skilful technical assistance. We thank the following organizations for funding the project: The Faculty of Health Sciences, University of Copenhagen (211-0610/09-3012); Dansk Kraeftforsknings fond; Kong Christian den Tiendes Fond; Civilingenioer Frode V. Nyegaard og Hustrus Fond and Harboefonden.