title: Mitochondrial dysfunction in choline deficiency-induced apoptosis in cultured rat hepatocytes.
abstract: Our recent studies have demonstrated that generation of ROS is associated with choline deficiency (CD)-induced apoptosis in CWSV-1 cells, an immortalized rat hepatocyte that becomes tumorigenic by stepwise culturing in decreasing levels of choline. In the present study, we investigated the effect of CD on loss of mitochondrial membrane potential (MMP), using the JC-1 probe by FASCAN assay. Our data demonstrate that MMP in CD-cultured cells was decreased in a time- and dose-dependent manner and that significant disruption occurred at 24 h, relative to high choline (HC, 70 microM) cultured cells. In order to investigate further the relationship among the CD-induced ROS, MMP collapse, and apoptosis, we examined the effects of different inhibitors on ROS production, MMP disruption, and apoptosis in CD or HC-cultured CWSV-1 cells. These data indicate that the disruption of MMP is an upstream event in CD-induced apoptosis, and mitochondrial dysfunction plays a key role in mediating CD-induced apoptosis in CWSV-1 cells.
authors.affiliation: Free Radical Biology & Aging Research Program, Oklahoma Medical Research Foundation, 825 NE 13th Street, MB21, Oklahoma City, OK 73104, USA.
authors.forename: Wei-Xing
authors.initials: WX
authors.lastname: Guo
authors.forename: Quentin N
authors.initials: QN
authors.lastname: Pye
authors.forename: Kelly S
authors.initials: KS
authors.lastname: Williamson
authors.forename: Charles A
authors.initials: CA
authors.lastname: Stewart
authors.forename: Kenneth L
authors.initials: KL
authors.lastname: Hensley
authors.forename: Yashige
authors.initials: Y
authors.lastname: Kotake
authors.forename: Robert A
authors.initials: RA
authors.lastname: Floyd
authors.forename: Robert H
authors.initials: RH
authors.lastname: Broyles
mesh_terms: D000818:Animals; D017209:Apoptosis; D001593:Benzyl Compounds / Q000494:pharmacology; D015153:Blotting, Western; D020169:Caspases / Q000378:metabolism; D002469:Cell Separation; D002478:Cells, Cultured; D002794:Choline / Q000378:metabolism*; D016572:Cyclosporine / Q000494:pharmacology; D053938:DNA Fragmentation; D004579:Electron Transport; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005434:Flow Cytometry; D005609:Free Radicals; D022781:Hepatocytes / Q000378:metabolism / Q000473:pathology*; D006845:Hydrocarbons, Fluorinated / Q000494:pharmacology; D008113:Liver Neoplasms / Q000378:metabolism; D008564:Membrane Potentials; D008928:Mitochondria / Q000378:metabolism / Q000473:pathology*; D051381:Rats; D017382:Reactive Oxygen Species; D012402:Rotenone / Q000494:pharmacology; D013997:Time Factors
chemical_list: D001593:Benzyl Compounds; C476029:Boc-D-FMK; D004791:Enzyme Inhibitors; D005609:Free Radicals; D006845:Hydrocarbons, Fluorinated; D017382:Reactive Oxygen Species; D012402:Rotenone; D016572:Cyclosporine; D020169:Caspases; D002794:Choline