title: Bcl-2 expression in synovial fibroblasts is essential for maintaining mitochondrial homeostasis and cell viability.
abstract: The regulation of proliferation and cell death is vital for homeostasis, but the mechanism that coordinately balances these events in rheumatoid arthritis (RA) remains largely unknown. In RA, the synovial lining thickens in part through increased proliferation and/or decreased synovial fibroblast cell death. Here we demonstrate that the anti-apoptotic protein, Bcl-2, is highly expressed in RA compared with osteoarthritis synovial tissues, particularly in the CD68-negative, fibroblast-like synoviocyte population. To determine the importance of endogenous Bcl-2, an adenoviral vector expressing a hammerhead ribozyme to Bcl-2 (Ad-Rbz-Bcl-2) mRNA was employed. Ad-Rbz-Bcl-2 infection resulted in reduced Bcl-2 expression and cell viability in synovial fibroblasts isolated from RA and osteoarthritis synovial tissues. In addition, Ad-Rbz-Bcl-2-induced mitochondrial permeability transition, cytochrome c release, activation of caspases 9 and 3, and DNA fragmentation. The general caspase inhibitor zVAD.fmk blocked caspase activation, poly(ADP-ribose) polymerase cleavage, and DNA fragmentation, but not loss of transmembrane potential or viability, indicating that cell death was independent of caspase activation. Ectopically expressed Bcl-xL inhibited Ad-Rbz-Bcl-2-induced mitochondrial permeability transition and apoptosis in Ad-Rbz-Bcl-2-transduced cells. Thus, forced down-regulation of Bcl-2 does not induce a compensatory mechanism to prevent loss of mitochondrial integrity and cell death in human fibroblasts.
authors.affiliation: Division of Rheumatology, Northwestern University Medical School, and Veterans Administration Chicago Healthcare System, Lakeside Division, Chicago, IL 60611, USA.
authors.forename: H
authors.initials: H
authors.lastname: Perlman
authors.forename: C
authors.initials: C
authors.lastname: Georganas
authors.forename: L J
authors.initials: LJ
authors.lastname: Pagliari
authors.forename: A E
authors.initials: AE
authors.lastname: Koch
authors.forename: K
authors.initials: K
authors.lastname: Haines
authors.forename: R M
authors.initials: RM
authors.lastname: Pope
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D017209:Apoptosis / Q000235:genetics; D001172:Arthritis, Rheumatoid / Q000378:metabolism / Q000473:pathology; D053148:Caspase 3; D061945:Caspase Inhibitors; D002452:Cell Count; D002470:Cell Survival / Q000235:genetics; D002478:Cells, Cultured; D015536:Down-Regulation / Q000235:genetics; D004789:Enzyme Activation; D005347:Fibroblasts / Q000378:metabolism* / Q000473:pathology / Q000502:physiology; D005822:Genetic Vectors / Q000494:pharmacology; D006706:Homeostasis / Q000235:genetics; D006801:Humans; D007425:Intracellular Membranes / Q000502:physiology; D008928:Mitochondria / Q000473:pathology / Q000502:physiology*; D010003:Osteoarthritis / Q000378:metabolism / Q000473:pathology; D010539:Permeability; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D016337:RNA, Catalytic / Q000235:genetics; D013583:Synovial Membrane / Q000378:metabolism* / Q000473:pathology / Q000502:physiology; D051020:bcl-X Protein
chemical_list: D000590:Amino Acid Chloromethyl Ketones; C494811:BCL2L1 protein, human; D061945:Caspase Inhibitors; D019253:Proto-Oncogene Proteins c-bcl-2; D016337:RNA, Catalytic; D051020:bcl-X Protein; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; C505407:CASP3 protein, human; D053148:Caspase 3