Induction of apoptosis in glioma cells by recombinant human Fas ligand.
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.


CONCLUSIONS
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.
Kawaguchi
S
S
Department of Neurosurgery, Saga Medical School, Japan.
Mineta
T
T
Ichinose
M
M
Masuoka
J
J
Shiraishi
T
T
Tabuchi
K
K
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001932:Brain Neoplasms / Q000473:pathology*; D002470:Cell Survival / Q000187:drug effects; D053222:Fas Ligand Protein; D005910:Glioma / Q000473:pathology*; D006801:Humans; D008562:Membrane Glycoproteins / Q000494:pharmacology*; D011817:Rabbits; D011994:Recombinant Proteins / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects* / Q000473:pathology
C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins