DAPT suppresses the proliferation of human glioma cell line SHG-44
Xin Liu1, Qiu-Ran Xu2, Wan-Fu Xie1
1Department of Neurosurgery, the First Affiliated Hospital of Medical College of Xi'an Jiaotong University, Xi'an, Shaanxi Province 710061, China.
Objective:
To explore the suppressing effect of γ-secretase inhibitor DAPT on proliferation of human glioma cell line SHG-44 in vitro and its mechanism.
Methods:
The SHG-44 cell was treated by DAPT with different concentration. The proliferation of cells was detected by MTT assay; cell cycle and TSC of CD133(+) were determined by flow cytometry analysis technique; the key factor in Notch signaling pathway (Notch-1, Delta-1, Hes-1) was measured by reverse transcriptase-polymerase chain reaction and western blotting.
Results:
DAPT inhibited the growth and proliferation of SHG-44 cells significantly(P<0.05). And the inhibiting effect on SHG-44 cells produced by DAPT showed a dose-dependent manner. DAPT increased the rate of cells in G0/G1 phase of SHG-44 cells, while it decreased the rate of cells in S phase. TSC of CD133(+) was significantly reduced after DAPT treated SHG-44 cells. The expression of protein and mRNA of Notch-1, Delta-1 and Hes-1 were gradually downregulated with the increase of DAPT doses.
Conclusions:
DAPT can downregulate these key factor in Notch signaling pathway, reduce the TSC of CD133+ and inhibit the proliferation of SHG-44 cells.
Insights
The γ-secretase inhibitor DAPT suppresses human glioma cell proliferation by downregulating Notch signaling pathway factors. This study reveals DAPT
Area of Science:
- Neuro-oncology
- Molecular Biology
- Cell Biology
Background:
- Glioma is a primary brain tumor with limited treatment options.
- The Notch signaling pathway plays a role in glioma cell proliferation and survival.
- γ-secretase inhibitors are being investigated for their anti-cancer properties.
Purpose of the Study:
- To investigate the inhibitory effect of DAPT on SHG-44 human glioma cells in vitro.
- To elucidate the mechanism underlying DAPT's effect on glioma cell proliferation.
- To examine DAPT's impact on the Notch signaling pathway in glioma cells.
Main Methods:
- SHG-44 cells were treated with varying concentrations of DAPT.
- Cell proliferation was assessed using MTT assays.
- Cell cycle distribution, CD133+ tumor stem cell (TSC) content, and Notch pathway markers (Notch-1, Delta-1, Hes-1) were analyzed via flow cytometry, RT-PCR, and Western blotting.
Main Results:
- DAPT significantly inhibited SHG-44 cell growth and proliferation in a dose-dependent manner.
- DAPT treatment led to an increase in G0/G1 phase cells and a decrease in S phase cells.
- DAPT reduced CD133+ TSC content and downregulated Notch-1, Delta-1, and Hes-1 at both mRNA and protein levels.
Conclusions:
- DAPT effectively inhibits human glioma cell proliferation in vitro.
- DAPT exerts its effect by downregulating key components of the Notch signaling pathway.
- DAPT reduces tumor stem cell content, suggesting potential therapeutic applications in glioma treatment.
