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Induction of apoptosis in glioma cell lines by TRAIL/Apo-2l
1Department of Molecular and Cell Biology, School of Life Sciences, University of Science and Technology of China, HeFei, China. wumian88@yahoo.com
Abstract:
TRAIL/Apo-2L, a novel cytokine, is a member of the tumor necrosis factor (TNF) family and serves as an extracellular signal triggering apoptosis. TRAIL/Apo-2L is capable of killing various transformed cells but not unstimulated primary T cells. In this study, five human glioma cells (U87, U118, U178, U563, and A172) were examined for their susceptibility to the apoptotic effects of TRAIL/Apo-2L. TRAIL/Apo-2L cDNA was isolated by RT-PCR, and recombinant TRAIL/Apo-2L protein was purified by the pMAL-c2 system (New England Biolabs, Beverly, MA). Exposure of A172 cells to bacterially expressed soluble TRAIL/Apo-2L fusion protein at a concentration of 1 microg/ml resulted in significant cell death over a 24-h period. Three experiments were performed to suggest that the TRAIL/Apo-2L killing was through the induction of apoptosis of A172 target cells. In addition, the expression of TRAIL/Apo-2L in different glioma cells was found to be variable, and TRAIL/Apo-2L-induced apoptosis was in a cell type-dependent manner. Some correlation between the susceptibility to TRAIL/Apo-2L and the expression level of one of its cognate receptors, DR4, was observed. In addition, cycloheximide worked synergistically with TRAIL/Apo-2L to induce apoptosis in glioma cells.
Insights
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/Apo-2L) induces apoptosis in human glioma cells. Its effectiveness varies by cell type and correlates with DR4 receptor expression, showing synergistic effects with cycloheximide.
Area of Science:
- Molecular Biology
- Cell Biology
- Oncology
Background:
- TRAIL/Apo-2L is a cytokine in the TNF family that triggers apoptosis.
- It effectively eliminates transformed cells while sparing normal T cells.
Purpose of the Study:
- To investigate the susceptibility of human glioma cells to TRAIL/Apo-2L-induced apoptosis.
- To explore the role of DR4 receptor expression and synergistic agents in TRAIL/Apo-2L-mediated cell death.
Main Methods:
- Isolation of TRAIL/Apo-2L cDNA via RT-PCR.
- Purification of recombinant TRAIL/Apo-2L protein.
- Exposure of A172 glioma cells to recombinant TRAIL/Apo-2L and assessment of cell death and apoptosis.
Main Results:
- Recombinant TRAIL/Apo-2L induced significant apoptosis in A172 glioma cells.
- TRAIL/Apo-2L expression and apoptosis induction varied among glioma cell lines.
- Susceptibility to TRAIL/Apo-2L showed correlation with DR4 receptor expression.
- Cycloheximide enhanced TRAIL/Apo-2L-induced apoptosis synergistically.
Conclusions:
- TRAIL/Apo-2L effectively induces apoptosis in human glioma cells in a cell type-dependent manner.
- DR4 receptor expression is a potential factor influencing glioma cell sensitivity to TRAIL/Apo-2L.
- Combined treatment with cycloheximide may enhance TRAIL/Apo-2L efficacy against glioma.