title: Expression of CD95(FAS) by gene transfer does not sensitize K562 to Fas-killing.
abstract: CD95(FAS/Apo-1) belongs to the family of TNF receptor related molecules and is expressed on many benign and malignant types of cells. CD95 triggering is involved in the apoptotic death of lymphoid cells and may also be important in myelopoiesis. Myeloid leukemia cells are in most cases resistant to CD95 triggering despite expressing the antigen. We reasoned that myeloid leukemic cells can become sensitized to the Fas pathway of cell death if CD95 expression is restored by gene transfer. As a model, we utilized the cell line K562 which does not express CD95 on their cell surface. The gene for CD95 was introduced into K562 cells (by electroporation). As a control, the gene coding for beta-galactosidase was used. The CD95 transfected K562 cells stably expressed CD95, and had a growth pattern comparable to untransfected cells, but still remained resistant to Fas-triggering (tested using recombinant Fas-ligand). Our experiment shows that antigen expression is not a critical determinant of Fas-mediated apoptosis, but further down-stream mechanisms determine the fate of the cells. The transfected cells also had a comparable susceptibility to NK-mediated lysis which shows that the expression of CD95 does not interfere with NK-mediated lysis of target cells.
authors.affiliation: Department of Molecular Hematology and Therapy, University of Texas, M.D. Anderson Cancer Center, Houston, USA.
authors.forename: R
authors.initials: R
authors.lastname: Munker
authors.forename: F
authors.initials: F
authors.lastname: Marini
authors.forename: S
authors.initials: S
authors.lastname: Jiang
authors.forename: C
authors.initials: C
authors.lastname: Savary
authors.forename: L
authors.initials: L
authors.lastname: Owen-Schaub
authors.forename: M
authors.initials: M
authors.lastname: Andreeff
mesh_terms: D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000235:genetics / Q000276:immunology; D018929:Cell Culture Techniques; D002455:Cell Division / Q000235:genetics / Q000276:immunology; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D019169:Jurkat Cells; D007694:Killer Cells, Natural / Q000276:immunology; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics* / Q000276:immunology* / Q000473:pathology; D014407:Tumor Cells, Cultured; D019014:fas Receptor / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology
chemical_list: D017304:Annexin A5; D019014:fas Receptor