Inhibition of Notch signaling pathway using γ-secretase inhibitor delivered by a low dose of Triton-X100 in cultured oral cancer cells
- Faculty of Dentistry, The University of Sydney, Sydney (Australia)
- School of Chemical & Biomolecular Engineering, The University of Sydney, Sydney (Australia)
Highlights: • A novel method is developed to deliver γ-secretase inhibitor DAPT using a low dose of Triton-X100. • This approach effectively blocks the cleavage of Notch1 receptor. • The elimination of Notch1 intracellular domain from cell nuclei is confirmed. • Notch1 signaling blockade in mRNA level is demonstrated upon applying this delivery system. How to effectively delivering therapeutic agents, including γ-secretase inhibitors (GSIs), into live cells, remains a significant challenge. This study assessed the effect of Notch signaling inhibition by examining levels of the Notch1 intracellular domain (N1ICD) in cultured oral cancer cells analyzed with random stitched images (2D) and 3D visualizations using confocal microscopy and quantitative gene analysis. Substantially, we have developed a novel method to assist the delivery of γ-secretase inhibitor, DAPT, into live cells in the presence of an effective minimum concentration of Triton-X100 (0.001%) without damaging cell activity and membrane integrity assessed with cell proliferation assays. The images obtained in this study showed that DAPT alone could not block the γ-secretase inhibitor despite inhibiting cell growth. Further analysis of quantitative gene expressions of Notch signaling canonical pathway to verify the effectiveness of the novel method for delivering inhibitor into live cells, displayed deregulation of Notch1, Delta-like ligand 1 (DLL1) and hairy and enhancer of split 1 (Hes1). Our data suggest that Notch1/Hes1 signaling pathway is deactivated using DAPT with a low dose of Triton-X100 in this cancer cells. And the finding also suggests that Notch1 could be engaged by DLL1 to promote differentiation in oral cancer cells. Using this approach, we demonstrate that Triton-X100 is a promising and effective permeabilization agent to deliver γ-secretase inhibitor DAPT into live oral epithelial cells. This strategy has the potential to implicate in the treatment of cancer diseases.
- OSTI ID:
- 23134362
- Journal Information:
- Biochemical and Biophysical Research Communications, Vol. 495, Issue 3; Other Information: Copyright (c) 2017 Elsevier Inc. All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); ISSN 0006-291X
- Country of Publication:
- United States
- Language:
- English
Similar Records
MAML1 regulates cell viability via the NF-{kappa}B pathway in cervical cancer cell lines
Dll4-Notch1 signaling but not VEGF-A is essential for hyperoxia induced vessel regression in retina