title: G-protein signaling abnormalities mediated by CD95 in salivary epithelial cells.
abstract: Salivary epithelial cells from patients with primary Sjögren's syndrome (SS) undergo Fas-mediated apoptosis. Bcl-2 and Bcl-xL are apoptosis suppressing oncogenes. Very little is known about the role of these oncogene molecules in salivary epithelial cells. To investigate the possible prevention of salivary glandular destruction in SS by Bcl-2 and Bcl-xL, stable transfectants expressing these molecules were made from HSY cells, a human salivary epithelial cell line. HSY cells were transfected with an expression vector for human Bcl-2 or Bcl-xL. Stable transfectants were selected and apoptosis was induced by anti-Fas antibody. Apoptosis was quantified by propidium iodide staining followed by flow cytometry. Caspase activity was detected by immunohistochemical analysis and enzyme cleavage of DEVD-AMC, a fluorescent substrate. Response to carbachol, a muscarinic receptor agonist, and EGF was measured by Ca2+ mobilization and influx. Fas-mediated apoptosis was significantly inhibited in Bcl-2 and Bcl-xL transfectants compared to wild-type and control transfectants (empty vector). Surprisingly, caspase activity was not inhibited in Bcl-2 and Bcl-xL transfectants. Activation of the Fas pathway in the Bcl-2 and Bcl-xL transfectants by antibody also inhibited carbachol and EGF responsiveness (i.e., Ca2+ mobilization and/or influx) by 50-60%. This Fas-mediated inhibition of cell activation was partially or completely restored by specific peptide interference of caspase enzyme activity. The prevention of Fas-mediated apoptosis by the overexpression of Bcl-2 and Bcl-xL in salivary gland epithelial cells results in injured cells expressing caspase activity and unable to respond normally to receptor agonists. Such damaged cells may exist in SS patients and could explain the severe dryness out of proportion to the actual number of apoptotic cells seen on salivary gland biopsy.
authors.affiliation: Department of Pediatrics, The University of Texas Health Science Center at San Antonio, San Antonio, Texas, TX 78229-3900, USA.
authors.forename: X B
authors.initials: XB
authors.lastname: Liu
authors.forename: R
authors.initials: R
authors.lastname: Masago
authors.forename: L
authors.initials: L
authors.lastname: Kong
authors.forename: B X
authors.initials: BX
authors.lastname: Zhang
authors.forename: S
authors.initials: S
authors.lastname: Masago
authors.forename: N
authors.initials: N
authors.lastname: Vela-Roch
authors.forename: M S
authors.initials: MS
authors.lastname: Katz
authors.forename: C K
authors.initials: CK
authors.lastname: Yeh
authors.forename: G H
authors.initials: GH
authors.lastname: Zhang
authors.forename: N
authors.initials: N
authors.lastname: Talal
authors.forename: H
authors.initials: H
authors.lastname: Dang
mesh_terms: D017209:Apoptosis; D002118:Calcium / Q000378:metabolism; D002217:Carbachol / Q000494:pharmacology; D002316:Cardiotonic Agents / Q000494:pharmacology; D020169:Caspases / Q000378:metabolism; D002518:Ceramides / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005434:Flow Cytometry; D019204:GTP-Binding Proteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism*; D012469:Salivary Glands / Q000166:cytology* / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D012859:Sjogren's Syndrome / Q000503:physiopathology*; D014162:Transfection; D014407:Tumor Cells, Cultured; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein; D019014:fas Receptor / Q000378:metabolism*
chemical_list: C494811:BCL2L1 protein, human; D002316:Cardiotonic Agents; D002518:Ceramides; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein; D019014:fas Receptor; D004815:Epidermal Growth Factor; D002217:Carbachol; D020169:Caspases; D019204:GTP-Binding Proteins; D002118:Calcium