Behavior of Nuclear Matrix Proteins during Camptothecin-Induced Apoptosis in HL-60 Human Leukemia Cells

Zweyer, Marina; Bareggi, Renato; Grill, Vittorio; Soranzo, Maria Rosa; Marugg, Richard A.; Riederer, Beat M.; Narducci, Paola; Martelli, Alberto M.

doi:10.1006/excr.1995.1349

Cited by 18 publications

(22 citation statements)

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“…Nucleolar remnants in apoptotic samples sometimes showed a large inner clear area ( Fig. 5D and E), as described for intact apoptotic HL-60 cells [Zweyer et al, 1995]. In this case, residual nucleoli presented also very electron-dense lenticular structures that were localized within the peripheral ring surrounding the internal electron-transparent area (Fig.…”

Section: Resultsmentioning

confidence: 81%

“…However, in the past, Arends et al [1990] ruled out biochemical and ultrastructural changes in the nuclear matrix prepared from apoptotic thymocytes. Nevertheless, indirect evidence suggesting a likely involvement of the nuclear matrix during apoptosis came from results obtained by means of immunofluorescence and/or immunoelectron microscopy techniques and showing unstainability of apoptotic nuclei with antibodies directed to nuclear matrix components as well as the presence in nuclei of apoptotic cells of granular and threaded bodies containing several nuclear matrix polypeptides [Miller et al, 1993;Zweyer et al, 1995;Weaver et al, 1996;Biggioggera 1997a,b;Zweyer et al, 1997;Lafarga et al, 1997]. However, firm evidence suggesting an involvement of the proteinaceous nuclear matrix during the apoptotic process was still missing.…”

Section: Discussionmentioning

confidence: 99%

“…However, the findings of Arends et al [1990] ruled out any differences both in the morphology and in the protein composition of nuclear matrix prepared from dexamethasone-treated thymocytes when compared to control. Subsequently, however, several articles have been published hinting at the fact that the nuclear matrix might be disassembled during the apoptotic process [Miller et al, 1993;Zweyer et al, 1995Zweyer et al, , 1997Tinnemans et al, 1995;Weaver et al, 1996]. It is thought that chromatin is organized in loops with an average size of 50-100 kb interacting with the nuclear matrix through specialized AT-rich DNA regions called SARs (scaffold-associated regions) or MARs (matrixassociated regions) [Boulikas, 1995;Bode et al, 1996].…”

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Biochemical and morphological characterization of the nuclear matrix from apoptotic HL-60 cells

et al. 1999

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We have characterized the nuclear matrix-intermediate filament fraction from control and apoptotic HL-60 cells. Apoptosis was induced by exposure to the topoisomerase I inhibitor, camptothecin. By means of two-dimensional polyacrylamide gel electrophoresis, striking qualitative and quantitative differences were seen in the protein composition of the nuclear matrix-intermediate filament fraction obtained from apoptotic cells in comparison with controls. Western blotting analysis of apoptotic nuclear matrix proteins revealed degradation of some (topoisomerase IIalpha, SAF-A) but not other (SATB1 and nucleolin) components. Moreover, immunofluorescent staining for typical matrix antigens (NuMA protein, lamin B, SC-35) showed that in 35-40% of the structures prepared from apoptotic samples, marked changes in the subnuclear distribution of these proteins were present. Striking morphological differences between control and apoptotic samples were also detected at the ultrastructural level. These results demonstrate that both biochemical and morphological changes can be detected in the nuclear matrix prepared from apoptotic HL-60 cells.

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Section: Resultsmentioning

confidence: 81%

Section: Discussionmentioning

confidence: 99%

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Biochemical and morphological characterization of the nuclear matrix from apoptotic HL-60 cells

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“…the 180-kDa nucleolar isoform of DNA topoisomerase II. 32 It has been reported by Zweyer 32 that the fluorescence patterns of the nucleolar isoform of topoisomerase II are well-maintained until the late phase of apoptosis in HL-60 human leukaemia cells.…”

Section: What Is the Functional Impact Of The Association Of The Nuclmentioning

confidence: 97%

Nucleolar segregation during apoptosis of haemopoietic stem cell line FDCP-Mix1

et al. 1999

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The programmed elimination of cells during apoptosis is distinct from necrosis both morphologically and biochemically. Currently, the morphological description of apoptosis discriminates between the segregation of the nucleolus and the so called`chromatin condensation'. The latter originates from observations of electron dense material adjacent to the nuclear envelope of apoptotic nuclei. Although there is ample evidence for an involvement of DNA in electron dense marginations, their true nature is still unknown. By studying apoptosis in FDCP-Mix, a pluripotent murine haemopoietic stem cell line, we found morphological and histochemical evidence that electron dense material at the nuclear envelope (NE) has emerged as a result of the segregation of nucleoli in association with the nuclear membrane. The remaining electron dense and homogenous bulk of the nucleolus labels for RNAse-gold, but even more intensely for DNAse-gold, and therefore could possibly be mistaken as`condensed chromatin' in the light microscope. The labelling of the electron dense material for DNase-gold in FDCP-Mix could be explained by a migration of DNA into the bulk of the nucleoli at an early stage of cell death.

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“…In addition, nucleolar ultrastructural changes that occur during apoptosis, such as nucleolar segregation and translocation of nucleolar proteins, are distinct from the changes observed during necrosis and may serve as an additional set of markers to differentiate between the two forms of cell death (Earnshaw 1995;Zweyer et al 1995Zweyer et al ,1997Biggiogera et al 1999;Martelli et al 2000).…”

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Nucleolar Changes and Fibrillarin Redistribution Following Apatone Treatment of Human Bladder Carcinoma Cells

Jamison

Gilloteaux²,

Perlaky

et al. 2010

J Histochem Cytochem.

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(KS) S U M M A R Y 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 ringshaped 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. (J Histochem Cytochem 58:635-651, 2010)

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Behavior of Nuclear Matrix Proteins during Camptothecin-Induced Apoptosis in HL-60 Human Leukemia Cells

Cited by 18 publications

References 0 publications

Biochemical and morphological characterization of the nuclear matrix from apoptotic HL-60 cells

Biochemical and morphological characterization of the nuclear matrix from apoptotic HL-60 cells

Nucleolar segregation during apoptosis of haemopoietic stem cell line FDCP-Mix1

Nucleolar Changes and Fibrillarin Redistribution Following Apatone Treatment of Human Bladder Carcinoma Cells

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