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Depletion of Bc1-2 by an antisense oligonucleotide induces apoptosis accompanied by oxidation and externalization of phosphatidylserine in NCI-H226 lung carcinoma cells

  • Chapter
Oxygen/Nitrogen Radicals: Cell Injury and Disease

Part of the book series: Developments in Molecular and Cellular Biochemistry ((DMCB,volume 37))

Abstract

Oxidant-induced apoptosis involves oxidation of many different and essential molecules including phospholipids. As a result of this non-specific oxidation, any signaling role of a particular phospholipid-class of molecules is difficult to elucidate. To determine whether preferential oxidation of phosphatidylserine (PS) is an early event in apoptotic signaling related to PS externalization and is independent of direct oxidant exposure, we chose a genetic-based induction of apoptosis. Apoptosis was induced in the lung cancer cell line NCI-H226 by decreasing the amount of Bc1-2 protein expression by preventing the translation ofbcl-2mRNA using an antisensebel-2oligonucleotide. Peroxidation of phospholipids was assayed using a fluorescent technique based on metabolic integration of an oxidation-sensitive and fluorescent fatty acid, cis-parinaric acid (PnA), into cellular phospholipids and subsequent HPLC separation of cis-PnA-labeled phospholipids. We found a decrease in Bc1-2 was associated with a selective oxidation of PS in a sub-population of the cells with externalized PS. No significant difference in oxidation of cis-PnA-labeled phospholipids was observed in cells treated with medium alone or a nonsense oligonucleotide. Treatment with either nonsense or antisensebcl-2oligonucleotides was not associated with changes in the pattern of individual phospholipid classes as determined by HPTLC. These metabolic and topographical changes in PS arrangement in plasma membrane appear to be early responses to antisensebcl-2exposure that trigger a PS-dependent apoptotic signaling pathway. This observed externalization of PS may facilitate the ‘labeling’ of apoptotic cells for recognition by macrophage scavenger receptors and subsequent phagocytic clearance. (Mol Cell Biochem 234/235: 125–133, 2002)

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Authors and Affiliations

  1. Department of Environmental and Occupational Health, University of Pittsburgh-Graduate School of Public Health, Pittsburgh, PA, USA

    Patrick P. Koty, Yulia Y. Tyurina, Shang-Xi Liu & Valerian E. Kagan

  2. Institute of Evolutionary Physiology and Biochemistry, St. Petersburg, Russia

    Yulia Y. Tyurina & Vladimir A. Tyurin

Authors
  1. Patrick P. Koty
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  2. Yulia Y. Tyurina
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  3. Vladimir A. Tyurin
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  4. Shang-Xi Liu
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  5. Valerian E. Kagan
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Editors and Affiliations

  1. Pathology and Physiology Research Branch, National Institute for Occupational Safety and Health, 1095 Willowdale Road Mall Stop B167, Morgantown, WV, 26505, USA

    Val Vallyathan , Xianglin Shi Ph.D.  & Vince Castranova ,  & 

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Koty, P.P., Tyurina, Y.Y., Tyurin, V.A., Liu, SX., Kagan, V.E. (2002). Depletion of Bc1-2 by an antisense oligonucleotide induces apoptosis accompanied by oxidation and externalization of phosphatidylserine in NCI-H226 lung carcinoma cells. In: Vallyathan, V., Shi, X., Castranova, V. (eds) Oxygen/Nitrogen Radicals: Cell Injury and Disease. Developments in Molecular and Cellular Biochemistry, vol 37. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-1087-1_14

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  • DOI: https://doi.org/10.1007/978-1-4615-1087-1_14

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-5388-1

  • Online ISBN: 978-1-4615-1087-1

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