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Apoptotic responsiveness of the Ewing's sarcoma family of tumours to tumour necrosis factor–related apoptosis‐inducing ligand (TRAIL)
Author(s) -
Van Valen Frans,
Fulda Simone,
Truckenbrod Borna,
Eckervogt Vera,
Sonnemann Jürgen,
Hillmann Axel,
Rödl Robert,
Hoffmann Christiane,
Winkelmann Winfried,
Schäfer Lutz,
DockhornDworniczak Barbara,
Wessel Torsten,
Boos Joachim,
Debatin KlausMichael,
Jürgens Herbert
Publication year - 2000
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/1097-0215(20001015)88:2<252::aid-ijc17>3.0.co;2-u
Subject(s) - apoptosis , programmed cell death , mg132 , biology , caspase , dna fragmentation , caspase 8 , cell culture , cytotoxic t cell , caspase 3 , microbiology and biotechnology , cycloheximide , poly adp ribose polymerase , necrosis , cancer research , proteasome inhibitor , biochemistry , polymerase , in vitro , enzyme , genetics
We investigated the cytotoxic responsiveness of 40 cell lines derived from representatives of the Ewing's sarcoma family of tumours (ESFT), i.e., Ewing's sarcoma (ES), peripheral primitive neuroectodermal tumour (pPNET) and Askin tumour (AT), to tumour necrosis factor–related apoptosis‐inducing ligand (TRAIL). Incubation with TRAIL at 100 ng/ml induced cell death at 24 hr in 19 of 26 ES, 11 of 12 pPNET and 2 of 2 AT cell lines. Half‐maximal cell death concentrations (IC 50 values) varied from 0.1 to 20 ng/ml. TRAIL displayed potent cytotoxic activity against freshly derived ESFT cell isolates. Cytotoxicity was associated with phosphatidylserine expression and internucleosomal DNA fragmentation, features characteristic of apoptosis. The apoptotic programme in the sensitive ESFT VH‐64 cell line revealed TRAIL‐induced activation of FLICE/MACH1 (caspase‐8) and CPP32/Yama/apopain (caspase‐3) and processing of the prototype caspase substrate poly(ADP‐ribose) polymerase. In addition, TRAIL provoked a collapse of the mitochondrial transmembrane potential (ΔΨ m ), parallelled by a reduction in ATP levels and release of cytochrome c from mitochondria into the cytosol. Inhibition of caspase‐8 and caspase‐3 by zIETDfmk and zDEVDfmk, respectively, substantially prevented TRAIL‐induced apoptosis. However, zIETDfmk, but not zDEVDfmk, reduced TRAIL‐mediated ΔΨ m dissipation, indicating that TRAIL causes mitochondrial dysfunction through caspase‐8 acting upstream of mitochondria. While macromolecule synthesis inhibitors (actinomycin D, cycloheximide) augmented susceptibility to TRAIL in TRAIL‐responsive cell lines, these agents did not render TRAIL‐resistant cell lines susceptible to TRAIL. However, the proteasome inhibitor MG132 sensitised to TRAIL in resistant cell lines. Collectively, these results show that TRAIL initiates effective death in the vast majority (80%) of cell lines derived from ESFT. Since TRAIL provoked cell death in ESFT ex vivo, this cytokine may be a promising drug for the treatment of ESFT in vivo. Int. J. Cancer 88:252–259, 2000. © 2000 Wiley‐Liss, Inc.