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Cell-death mechanisms in the IPLB-LdFB insect cell line: a nuclear located Bcl-2-like molecule as a possible controller of 2-deoxy-D-ribose-mediated DNA fragmentation

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Abstract

In the IPLB-LdFB insect cell line, oncosis and apoptosis are the two pre-mortal processes, whereas necrosis is the post-mortem condition. As found in mammals, adenosine triphosphate depletion of insect cells by oligomycin A induces oncosis. The apoptotic inducer 2-deoxy-D-ribose (dRib) provokes cell death through an intrinsic apoptotic pathway similar to that observed in mammalian models and results in oligonucleosomal DNA fragmentation. The addition to insect cells of an anti-Bcl-2 polyclonal antibody known to prevent dRib-mediated apoptosis abolishes DNA fragmentation, whereas cytochrome c release and the increase in a caspase 3-like activity are still detectable. These and previous findings suggest a double role for the Bcl-2-like molecule in IPLB-LdFB, i.e. the maintenance of mitochondrial integrity and the control of apoptotic machinery at the nuclear level.

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References

  • Alnemri ES, Robertson NM, Fernandes TF, Croce CM, Litwack G (1992) Overexpressed full-length human BCL2 extends the survival of baculo-virus-infected Sf9 insect cells. Proc Natl Acad Sci USA 89:7295–7299

    Google Scholar 

  • Barbieri D, Malagoli D, Cuoghi B, Ottaviani E (2001) An anti-Bcl-2 antibody prevents 2-deoxy-D-ribose-induced apoptosis in the IPLB-LdFB insect cell line. Cell Mol Life Sci 58:653–659

    Google Scholar 

  • Blackstone NW, Green DR (1999) The evolution of a mechanism of cell suicide. BioEssays 21:84–88

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Chen-Levy Z, Nourse J, Clearly ML (1989) The bcl-2 candidate proto-oncogene product is a 24-kilodalton integral-membrane protein highly expressed in lymphoid cell lines and lymphomas carrying the t(14;18) translocation. Mol Cell Biol 9:701–710

    Google Scholar 

  • Colussi PA, Quinn LM, Huang DCS, Coombe M, Read SH, Richardson H, Kumar S (2000) Debcl, a proapoptotic Bcl-2 homologue, is a component of Drosophila melanogaster cell death machinery. J Cell Biol 148:703–714

    Google Scholar 

  • Dartsch DC, Schaefer A, Boldt S, Kolch W, Marquardt H (2002) Comparison of anthracycline-induced death of human leukemia cells: programmed cell death versus necrosis. Apoptosis 7:537–548

    Google Scholar 

  • Eguchi Y, Shimizu S, Tsujimoto Y (1997) Intracellular ATP levels determine cell death fate by apoptosis or necrosis. Cancer Res 57:1835–1840

    CAS  PubMed  Google Scholar 

  • Ferri KF, Kroemer G (2001) Mitochondria—the suicide organelles. BioEssays 23:111–115

    Google Scholar 

  • Gross A, McDonnell JM, Korsmeyer SJ (1999) BCL-2 family members and the mitochondria in apoptosis. Genes Dev 13:1899–1911

    CAS  PubMed  Google Scholar 

  • Hockenbery DM, Olivai ZN, Yin XM, Milliman CL, Korsmeyer SJ (1993) Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell 75:241–251

    Article  Google Scholar 

  • Jacobson MD, Burne JF, King MD, Miyashita T, Reed JC, Raff MC (1993) Bcl-2 blocks apoptosis in cell lacking mitochondrial DNA. Nature 361:365–368

    Google Scholar 

  • Kletsas D, Barbieri D, Stathakos D, Botti B, Bergamini S, Tomasi A, Monti D, Malorni W, Franceschi C (1998) The highly reducing sugar 2-deoxy-D-ribose induces apoptosis in human fibroblasts by reduced glutathione depletion and cytoskeletal disruption. Biochem Biophys Res Commun 243:416–425

    Google Scholar 

  • Krajewski S, Tamaka S, Schibler MJ, Fenton W, Reed JC (1993) Investigation of the subcellular distribution of the bcl-2 oncoprotein: residence in the nuclear envelope, endoplasmic reticulum, and outer mitochondrial membranes. Cancer Res 53:4710–4714

    Google Scholar 

  • Lynn DE, Dougherty EM, McClintock JT, Loeb M (1988) Development of cell lines from various tissues of Lepidoptera. In: Kuroda Y, Kurstak E, Maramorosch K (eds) Invertebrate and fish tissue culture. Japan Scientific Societies, Tokyo, pp 239–242

    Google Scholar 

  • Majno G, Joris I (1995) Apoptosis, oncosis, and necrosis. An overview of cell death. Am J Pathol 146:3–15

    CAS  PubMed  Google Scholar 

  • Mayer M, Noble M (1994) N-acetyl-L-cysteine is a pluripotent protector against cell death and enhancer of trophic factor-mediated cell survival in vitro. Proc Natl Acad Sci USA 91:7496–7500

    Google Scholar 

  • McConkey DJ (1998) Biochemical determinants of apoptosis and necrosis. Toxicol Lett 99:157–168

    Google Scholar 

  • Monaghan P, Robertson D, Amos TAS, Dyer MJS, Mason DY, Greaves MF (1992) Ultrastructural localization of BCL-2 protein. J Histochem Cytochem 40:1819–1825

    Google Scholar 

  • Nagata S (2000) Apoptotic DNA fragmentation. Exp Cell Res 256:12–18

    Google Scholar 

  • Ottaviani E, Barbieri D, Franchini A, Kletsas D (2000) PDGF and TGF-β partially prevent 2-deoxy-D-ribose-induced apoptosis in the fat body cell line IPLB-LdFB from the insect Lymantria dispar. J Insect Physiol 46:81–87

    Google Scholar 

  • Ottaviani E, Barbieri D, Malagoli D, Kletsas D (2001) Involvement of PI 3-kinase, PKA and PKC in PDGF- and TGF-β-mediated prevention of 2-deoxy-D-ribose-induced apoptosis in the insect cell line, IPLB-LdFB. Cell Biol Int 25:171–177

    Google Scholar 

  • Quinn LM, Coombe M, Mills K, Daish T, Colussi PA, Kumar S, Richardson H (2003) Buffy, a Drosophila Bcl-2 protein, has anti-apoptotic and cell cycle inhibitory functions. EMBO J 22:3568–3579

    Google Scholar 

  • Richardson H, Kumar S (2002) Death to flies: Drosophila as a model system to study programmed cell death. J Immunol Methods 265:21–38

    Google Scholar 

  • Robertson JD, Orrenius S, Zhivotovsky B (2000) Review: nuclear events in apoptosis. J Struct Biol 129:346–358

    Google Scholar 

  • Shimizu S, Eguchi Y, Kamiike W, Itoh Y, Hasegawa J, Yamabe K, Otsuki Y, Matsuda H, Tsujimoto Y (1996a) Induction of apoptosis as well as necrosis by hypoxia and predominant prevention of apoptosis by Bcl-2 and Bcl-XL. Cancer Res 56:2161–2166

    Google Scholar 

  • Shimizu S, Eguchi Y, Kamiike W, Waguri S, Uchiyama Y, Matsuda H, Tsujimoto Y (1996b) Retardation of chemical hypoxia-induced necrotic cell death by Bcl-2 and ICE inhibitors: possible involvement of common mediators in apoptotic and necrotic signal transductions. Oncogene 12:2045–2050

    Google Scholar 

  • Voehringer DW, McConkey DJ, McDonnel TJ, Brisbay S, Meyn RE (1998) Bcl-2 expression causes redistribution of glutathione to the nucleus. Proc Natl Acad Sci USA 95:2956–2960

    Google Scholar 

  • Wu YC, Stanfield GM, Horvitz HR (2000) NUC-1, a Caenorhabditis elegans DNase II homolog, functions in an intermediate step of DNA degradation during apoptosis. Gene Dev 14:536–548

    Google Scholar 

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Acknowledgements

The authors are grateful to Dr. D. E. Lynn (USDA/ARS, Insect Biocontrol Laboratory, BARC-West, Beltsville, Md., USA) who kindly supplied the cell line.

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Correspondence to Enzo Ottaviani.

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This work was supported by a MIUR (Italy) grant to E.O.

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Malagoli, D., Iacconi, I., Marchesini, E. et al. Cell-death mechanisms in the IPLB-LdFB insect cell line: a nuclear located Bcl-2-like molecule as a possible controller of 2-deoxy-D-ribose-mediated DNA fragmentation. Cell Tissue Res 320, 337–343 (2005). https://doi.org/10.1007/s00441-004-1050-z

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  • DOI: https://doi.org/10.1007/s00441-004-1050-z

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