title: Caspase 8 activation independent of Fas (CD95/APO-1) signaling may mediate killing of B-chronic lymphocytic leukemia cells by cytotoxic drugs or gamma radiation.
abstract: Ligation of the cell-surface Fas molecule by its ligand (Fas-L) or agonistic anti-Fas monoclonal antibodies results in the cleavage and activation of the cysteine protease procaspase 8 followed by the activation of procaspase 3 and by apoptosis. In some leukemia cell lines, cytotoxic drugs induce expression of Fas-L, which may contribute to cell killing through the ligation of Fas. The involvement of Fas, Fas-L, and caspase 8 was studied in the killing of B-cell chronic lymphocytic leukemia (B-CLL) cells by chlorambucil, fludarabine, or gamma radiation. Spontaneous apoptosis was observed at 24-hour incubation, with additional apoptosis induced by each of the cytotoxic treatments. Although Fas mRNA expression was elevated after exposure to chlorambucil, fludarabine, or gamma radiation, Fas protein levels only increased after irradiation. Therefore, Fas expression may be regulated by multiple mechanisms that allow the translation of Fas mRNA only in response to restricted cytotoxic stimuli. None of the cytotoxic stimuli studied here induced Fas-L expression. An agonistic anti-Fas monoclonal antibody (CH-11) did not significantly augment apoptosis induction by any of the death stimuli. A Fas-blocking antibody (ZB4) did not inhibit spontaneous, chlorambucil-, fludarabine-, or radiation-induced apoptosis. However, procaspase 8 processing was induced by all cytotoxic stimuli. These data suggest that the Fas/Fas-L signaling system does not play a major role in the induction of apoptosis in B-CLL cells treated with cytotoxic drugs or radiation. However, Fas-independent activation of caspase 8 may play a crucial role in the regulation of apoptosis in these cells.
authors.affiliation: Department of Hematology, Royal Free and University College Medical School, Royal Free Campus, London, United Kingdom.
authors.forename: D T
authors.initials: DT
authors.lastname: Jones
authors.forename: K
authors.initials: K
authors.lastname: Ganeshaguru
authors.forename: A E
authors.initials: AE
authors.lastname: Virchis
authors.forename: N I
authors.initials: NI
authors.lastname: Folarin
authors.forename: M W
authors.initials: MW
authors.lastname: Lowdell
authors.forename: A B
authors.initials: AB
authors.lastname: Mehta
authors.forename: H G
authors.initials: HG
authors.lastname: Prentice
authors.forename: A V
authors.initials: AV
authors.lastname: Hoffbrand
authors.forename: R G
authors.initials: RG
authors.lastname: Wickremasinghe
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D001402:B-Lymphocytes / Q000187:drug effects / Q000473:pathology / Q000528:radiation effects; D053181:Caspase 8; D053453:Caspase 9; D020169:Caspases / Q000378:metabolism / Q000494:pharmacology*; D004347:Drug Interactions; D004789:Enzyme Activation; D053222:Fas Ligand Protein; D005260:Female; D005720:Gamma Rays; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000188:drug therapy / Q000473:pathology* / Q000532:radiotherapy; D008297:Male; D008562:Membrane Glycoproteins / Q000032:analysis; D015398:Signal Transduction; D019014:fas Receptor / Q000032:analysis / Q000276:immunology / Q000494:pharmacology*
chemical_list: D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D019014:fas Receptor; C505530:CASP8 protein, human; C506433:CASP9 protein, human; D053181:Caspase 8; D053453:Caspase 9; D020169:Caspases