Irradiation induced clonogenic cell death of human malignant glioma cells does not require CD95/CD95L interactions.
BACKGROUND
Radiotherapy is the single most effective therapy for malignant gliomas. Targeting the CD95 apoptotic pathway is a promising experimental approach to these neoplasms. Here, we asked whether irradiation modulates CD95-mediated apoptosis of human malignant glioma cells in vitro.


METHODS
LN-18, LN-229 and T98G human malignant glioma cell lines were irradiated with dosages from 0-8 Gy and treated with CD95L (CD95 ligand). CD95 expression was assessed by flow cytometry. Caspase activity was determined by DEVD cleavage. Cytotoxic effects were assessed by crystal violet staining of cells in a 96-well plate assay. Clonogenic cell death was determined by a standard colony forming assay.


RESULTS
We find that (i) CD95L-induced apoptosis, but not irradiation-induced clonogenic cell death, involves caspase 3 activation and is blocked by the viral caspase inhibitor, crm-A. (ii) Irradiation does not modulate CD95 expression either in p53 wild-type or in p53 mutant glioma cell lines, and does not enhance CD95L-evoked caspase 3 activity or CD95L-induced clonogenic cell death.


CONCLUSIONS
We conclude that endogenous CD95/CD95L interactions are not involved in radiation-induced clonogenic cell death and that the killing cascades of CD95L and irradiation are independent in human malignant glioma cells.
Streffer
J R
JR
Department of Neurology, University of Tübingen, Germany.
Schuster
M
M
Pohl
U
U
Belka
C
C
Dichgans
J
J
Bamberg
M
M
Weller
M
M
D017209:Apoptosis / Q000528:radiation effects*; D001932:Brain Neoplasms / Q000276:immunology / Q000473:pathology*; D053222:Fas Ligand Protein; D005910:Glioma / Q000276:immunology / Q000473:pathology*; D006801:Humans; D008562:Membrane Glycoproteins / Q000276:immunology*; D019014:fas Receptor / Q000276:immunology*
C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D019014:fas Receptor