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Induction of apoptosis in glioma cells by recombinant human Fas ligand
S Kawaguchi1, T Mineta, M Ichinose
1Department of Neurosurgery, Saga Medical School, Japan.
Objective:
Fas ligand (FasL) belongs to the tumor necrosis factor family and has the ability to induce apoptosis in susceptible target cells by binding to its receptor, Fas. It has been demonstrated recently that the FasL/Fas system plays a pivotal role in the cytocidal activity of T lymphocytes in the immune system. FasL may act as a cytotoxic effector molecule to Fas-expressing malignant tumor cells. We reported previously that Fas is commonly expressed in human brain tumor cells. In this study, we examine the possible application of FasL to therapy for malignant brain tumors.
Methods:
To develop an expression system yielding large amounts of FasL, we constructed a baculovirus vector containing complementary deoxyribonucleic acid of human FasL under the control of a polyhedrin promoter. We produced human FasL in Spodoptera frugiperda (Sf9) insect cells infected by the recombinant baculovirus carrying FasL complementary deoxyribonucleic acid and studied the cytocidal activity of FasL against the T98G human glioblastoma cell line.
Results:
FasL expression in Sf9 cells was confirmed immunocytochemically with rabbit antibody raised against the cytoplasmic domain of human FasL. The FasL released into the supernatant of cultured Sf9 cells was also verified by Western blotting, and it specifically induced apoptosis in T98G cells. The induced apoptosis by recombinant human FasL was confirmed by annexin V-fluorescein isothiocyanate staining.
Conclusion:
The present results suggest that the induction of apoptosis by the Fas/FasL system could be a new strategy for the treatment of malignant brain tumors, which are resistant to conventional therapies.
Insights
Fas ligand (FasL) induces apoptosis in malignant brain tumor cells. This Fas/FasL system offers a novel therapeutic strategy for brain tumors resistant to conventional treatments.
Area of Science:
- Molecular Biology
- Immunology
- Oncology
Background:
- Fas ligand (FasL) is a tumor necrosis factor family member inducing apoptosis.
- The FasL/Fas system is crucial for T lymphocyte cytotoxic activity.
- FasL may target Fas-expressing malignant tumor cells, including brain tumors.
Purpose of the Study:
- To investigate the therapeutic potential of FasL for malignant brain tumors.
- To develop an efficient system for producing human FasL.
Main Methods:
- Constructed a baculovirus vector for human FasL expression.
- Produced recombinant human FasL in Sf9 insect cells.
- Assessed FasL's cytocidal activity against T98G glioblastoma cells.
Main Results:
- Confirmed FasL expression and release from Sf9 cells.
- Demonstrated that recombinant human FasL specifically induces apoptosis in T98G cells.
- Verified apoptosis induction using annexin V-fluorescein isothiocyanate staining.
Conclusions:
- The Fas/FasL system represents a promising new strategy for treating malignant brain tumors.
- This approach could overcome resistance to conventional therapies.
