title: Nucleolar changes and fibrillarin redistribution following apatone treatment of human bladder carcinoma cells.
abstract: Ascorbate and menadione (Apatone) in a ratio of 100:1 kills tumor cells by autoschizis. In this study, vitamin-induced changes in nucleolar structure were evaluated as markers of autoschizis. Human bladder carcinoma (T24) cells were overlain with vitamins or with culture medium. Supernatants were removed at 1-hr intervals from 1 to 4 hr, and the cells were washed with PBS and prepared for assay. Apatone produced marked alterations in nucleolar structure including redistribution of nucleolar components, formation of ring-shaped nucleoli, condensation and increase of the proportion of perinucleolar chromatin, and the enlargement of nucleolar fibrillar centers. Immunogold labeling of the nucleolar rRNA revealed a granular localization in treated and sham-treated cells, and immunogold labeling of the rDNA revealed a shift from the fibrillar centers to the condensed perinucleolar chromatin. Fibrillarin staining shifted from the fibrillar centers and adjacent regions to a more homogeneous staining of the entire nucleolus and was consistent with the percentage of autoschizic cells detected by flow cytometry. Because autoschizis entails sequential reactivation of DNase I and DNase II, and because the fibrillarin redistribution following DNase I and Apatone treatment is identical, it appears that the nucleolar and fibrillarin changes are markers of autoschizis.
authors.affiliation: St. Thomas Hospital, Akron, Ohio 44310, USA. jamisonj@summa-health.org
authors.forename: James M
authors.initials: JM
authors.lastname: Jamison
authors.forename: Jacques
authors.initials: J
authors.lastname: Gilloteaux
authors.forename: Laszlo
authors.initials: L
authors.lastname: Perlaky
authors.forename: Marc
authors.initials: M
authors.lastname: Thiry
authors.forename: Karel
authors.initials: K
authors.lastname: Smetana
authors.forename: Deborah
authors.initials: D
authors.lastname: Neal
authors.forename: Karen
authors.initials: K
authors.lastname: McGuire
authors.forename: Jack L
authors.initials: JL
authors.lastname: Summers
mesh_terms: D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology*; D001205:Ascorbic Acid / Q000494:pharmacology*; D002277:Carcinoma / Q000737:chemistry / Q000648:ultrastructure*; D045744:Cell Line, Tumor; D002466:Cell Nucleolus / Q000737:chemistry / Q000187:drug effects / Q000648:ultrastructure*; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis*; D006801:Humans; D001749:Urinary Bladder Neoplasms / Q000737:chemistry / Q000648:ultrastructure*; D014812:Vitamin K / Q000494:pharmacology; D024483:Vitamin K 3 / Q000494:pharmacology*
chemical_list: D002868:Chromosomal Proteins, Non-Histone; C047458:fibrillarin; D014812:Vitamin K; D024483:Vitamin K 3; D001205:Ascorbic Acid
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