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Pyrrolidine dithiocarbamate (PDTC) blocks apoptosis and promotes ionizing radiation-induced necrosis of freshly-isolated normal mouse spleen cells

Apoptosis volume 15, pages 705–714 (2010)Cite this article

Abstract

Ionizing radiation (IR) is a pro-oxidant that kills cells by both apoptotic and necrotic mechanisms. Pyrrolidine dithiocarbamate (PDTC) is a thiol-containing compound that may act either as a pro- or anti-oxidant depending on the experimental conditions. This study was designed to determine whether PDTC would reduce or enhance IR-induced cell death of freshly-isolated normal mouse B6/129 spleen cells (NMSC). We determined the effect of increasing doses of IR, PDTC alone and PDTC followed by IR on the viability of NMSC. Annexin V and propidium iodide (Annexin V/PI) staining demonstrated a dose and time-dependent relationship in which PDTC enhanced the percentage of IR-induced apoptotic/necrotic NMSC. Trypan blue dye inclusion confirmed that a loss of membrane integrity was occurring 1 h after incubation with PDTC plus IR. Reduction in the glutathione (GSH)/glutathione disulfide (GSSG) ratio and GSH demonstrated that both IR (8.5 Gy) and PDTC acted as pro-oxidants, but their mechanisms of action differed: In contrast to IR, which promoted p53 activation and caspase 3/7-mediated apoptosis, PDTC inhibited IR-induced p53 and caspase 3/7 activity. However, PDTC increased H2O2 formation and necrosis, resulting in an overall increase in IR-induced cell death. Catalase prevented the PDTC-induced increase in IR cytotoxicity implicating the generation of H2O2 as a major factor in this mechanism. These results demonstrate that in NMSC PDTC acts as pro-oxidant and enhances IR-induced cell cytotoxicity by increasing H2O2formation and thiol oxidation. As such, they strongly suggest that the use of PDTC as an adjunct to reduce radiation toxicity should be avoided.

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Acknowledgment

Supported by Merit Review and Type II Merit Review Grants from the Veterans Affairs Administration and by HL-709.

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Affiliations

  1. Veterans Affairs Medical Center, Lexington, KY, 40511, USA

    John S. Thompson, Andrea A. Tapp, Brandy Nelson, Yanxia Chu, Judith Glass, Micheal Moneyhon & Stephen A. Brown

  2. Department of Internal Medicine, University of Kentucky, Lexington, KY, 40536, USA

    John S. Thompson, Andrea A. Tapp, Brandy Nelson, Yanxia Chu, Judith Glass, Micheal Moneyhon & Stephen A. Brown

  3. Department of Internal Medicine, University of Kentucky, 800 Rose Street, Lexington, KY, 40536, USA

    John S. Thompson

  4. The Office of the Dean, School of Health Professions, University of Texas Health Science Center at San Antonio, San Antonio, TX, 78229, USA

    Reto Asmis

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  1. John S. Thompson
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  2. Reto Asmis
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Correspondence to John S. Thompson.

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Thompson, J.S., Asmis, R., Tapp, A.A. et al. Pyrrolidine dithiocarbamate (PDTC) blocks apoptosis and promotes ionizing radiation-induced necrosis of freshly-isolated normal mouse spleen cells. Apoptosis 15, 705–714 (2010). https://doi.org/10.1007/s10495-010-0487-7

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  • DOI: https://doi.org/10.1007/s10495-010-0487-7

Keywords

  • Ionizing radiation
  • Pyrrolidine dithiocarbamate
  • Caspase
  • Hydrogen peroxide
  • Catalase