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

  • Patrick P. Koty
  • Yulia Y. Tyurina
  • Vladimir A. Tyurin
  • Shang-Xi Liu
  • Valerian E. Kagan
Part of the Developments in Molecular and Cellular Biochemistry book series (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)

Key words

antisense Bcl-2 apoptosis phosphatidylserine oxidation externalization 

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Copyright information

© Springer Science+Business Media New York 2002

Authors and Affiliations

  • Patrick P. Koty
    • 1
  • Yulia Y. Tyurina
    • 1
    • 2
  • Vladimir A. Tyurin
    • 2
  • Shang-Xi Liu
    • 1
  • Valerian E. Kagan
    • 1
  1. 1.Department of Environmental and Occupational HealthUniversity of Pittsburgh-Graduate School of Public HealthPittsburghUSA
  2. 2.Institute of Evolutionary Physiology and BiochemistrySt. PetersburgRussia

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