Fas ligand is present in tumors of the Ewing's sarcoma family and is cleaved into a soluble form by a metalloproteinase.
Fas ligand (FasL) exists in transmembrane and soluble forms and induces apoptosis on cross-linking with the Fas receptor. We evaluated the biological significance of FasL and Fas in 61 tumor tissues and 9 cell lines of the Ewing's sarcoma family of tumors (ESFT). FasL was present in 62.5% and Fas in 79.4% of primary ESFT. Metastatic tumors had higher expression of FasL (95%), suggesting association with a metastatic phenotype. FasL was detected in the cytoplasm and membrane of ESFT cells by immunofluorescence. Western blotting revealed transmembrane and soluble FasL in cytosolic extracts and soluble FasL in conditioned media. Both transmembrane and soluble FasL induced apoptosis of Fas-sensitive Jurkat cells in co-culture experiments with ESFT cells or their media. Treatment with phenanthroline and the synthetic metalloproteinase inhibitor BB-3103 reduced the levels of soluble FasL in the media, suggesting that in ESFT, FasL is processed by a metalloproteinase and released in the extracellular milieu. The released soluble FasL may serve to attack cells of the immune system and/or interfere with the binding of transmembrane FasL with Fas, and results in down-regulation of transmembrane FasL. Synthetic metalloproteinase inhibitors may modify the ratio of transmembrane to soluble FasL.
Mitsiades
N
N
Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA. nmitsiad@box-n.nih.gov
Poulaki
V
V
Kotoula
V
V
Leone
A
A
Tsokos
M
M
D000906:Antibodies / Q000494:pharmacology; D017209:Apoptosis; D002462:Cell Membrane / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D007150:Immunohistochemistry; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors; D009362:Neoplasm Metastasis; D010618:Phenanthrolines / Q000494:pharmacology; D012512:Sarcoma, Ewing / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000494:pharmacology; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D019014:fas Receptor / Q000276:immunology / Q000378:metabolism* / Q000502:physiology
D000906:Antibodies; C108282:BB 3103; D017077:Culture Media, Conditioned; D006877:Hydroxamic Acids; D010618:Phenanthrolines; D019715:Tissue Inhibitor of Metalloproteinase-1; D019014:fas Receptor; D019716:Tissue Inhibitor of Metalloproteinase-2; D008666:Metalloendopeptidases; C025205:1,10-phenanthroline