Elsevier

Experimental Cell Research

Volume 279, Issue 1, 10 September 2002, Pages 153-165
Experimental Cell Research

Regular Article
A Role for the de Novo Sphingolipids in Apoptosis of Photosensitized Cells

https://doi.org/10.1006/excr.2002.5599Get rights and content

Abstract

Sphingolipids have been implicated in apoptosis after various stress inducers. To assess the involvement of the de novo sphingolipid pathway in apoptosis, photodynamic therapy (PDT) with the photosensitizer Pc 4 was used as a novel stress inducer. Here we provide biochemical and genetic evidence of the role of the de novo sphingolipids in apoptosis post-Pc 4-PDT. In Jurkat cells PDT-induced intracellular sphinganine accumulation, DEVDase activation, PARP cleavage, and apoptosis were suppressed by the de novo sphingolipid synthesis inhibitor ISP-1 (Myriocin). Coincubation with sphinganine, sphingosine, or C16-ceramide specifically reversed the antiapoptotic actions of ISP-1 or the singlet oxygen scavenger l-histidine. PDT-induced cytochrome c release from mitochondria into the cytosol was inhibited by l-histidine, but not by ISP-1. Cotreatment with sphinganine did not reverse the inhibitory effect of l-histidine. In addition, PDT-induced sphinganine accumulation and apoptosis were ISP-1-sensitive in A431 human epidermoid and HT29 human carcinoma cells. Furthermore, in LY-B cells, CHO-derived mutants deficient in the de novo sphingolipid synthesis enzyme serine palmitoyltransferase (SPT) activity, DEVDase activation and apoptosis were delayed and suppressed post-PDT. Hence, the data are consistent with the partial involvement of the de novo sphingolipid pathway in apoptosis via DEVDase activation downstream of mitochondrial cytochrome c release post-Pc 4-PDT.

References (68)

  • T. Herget et al.

    Production of ceramides causes apoptosis during early neural differentiation in vitro

    J. Biol. Chem.

    (2000)
  • D.K. Perry et al.

    Serine palmitoyltransferase regulates de novo ceramide generation during etoposide-induced apoptosis

    J. Biol. Chem.

    (2000)
  • Y. Miyake et al.

    Serine palmitoyltransferase is the primary target of a sphingosine-like immunosuppressant, ISP-1/myriocin

    Biochem. Biophys. Res. Commun.

    (1995)
  • K. Hanada et al.

    Specificity of inhibitors of serine palmitoyltransferase (SPT), a key enzyme in sphingolipid biosynthesis, in intact cells: A novel evaluation system using an SPT-defective mammalian cell mutant

    Biochem. Pharmacol.

    (2000)
  • B. Nagy et al.

    Fumonisin B1 does not prevent apoptosis in A431 human epidermoid carcinoma cells after photosensitization with phthalocyanine 4

    J. Photochem. Photobiol. B.

    (2000)
  • B. Nagy et al.

    FADD null mouse embryonic fibroblasts undergo apoptosis after photosensitization with the silicon phthalocyanine Pc 4

    Arch. Biochem. Biophys.

    (2001)
  • P. Santana et al.

    Acid sphingomyelinase-deficient human lymphoblasts and mice are defective in radiation-induced apoptosis

    Cell

    (1996)
  • A.H. Merrill et al.

    Quantitation of free sphingosine in liver by high-performance liquid chromatography

    Anal. Biochem.

    (1988)
  • M.E. Varnes et al.

    Photodynamic therapy-induced apoptosis in lymphoma cells: Translocation of cytochrome c causes inhibition of respiration as well as caspase activation

    Biochem. Biophys. Res. Commun.

    (1999)
  • J. Chaudiere et al.

    Intracellular antioxidants: from chemical to biochemical mechanisms

    Food Chem. Toxicol.

    (1999)
  • I. Belichenko et al.

    Caspase-resistant vimentin suppresses apoptosis after photodynamic treatment with a silicon phthalocyanine in Jurkat cells

    Arch. Biochem. Biophys.

    (2001)
  • I.E. Kochevar et al.

    Photosensitized production of singlet oxygen

    Methods Enzymol.

    (2000)
  • L.Y. Xue et al.

    Photodynamic therapy-induced death of MCF-7 human breast cancer cells: a role for caspase-3 in the late steps of apoptosis but not for the critical lethal event

    Exp. Cell Res.

    (2001)
  • K. Hanada et al.

    Mammalian cell mutants resistant to a sphingomyelin-directed cytolysin: Genetic and biochemical evidence for complex formation of the LCB1 protein with the LCB2 protein for serine palmitoyltransferase

    J. Biol. Chem.

    (1998)
  • G.M. Jenkins et al.

    Role for de novo sphingoid base biosynthesis in the heat-induced transient cell cycle arrest of Saccharomyces cerevisiae

    J. Biol. Chem.

    (2001)
  • L.L. Listenberger et al.

    Palmitate-induced apoptosis can occur through a ceramide-independent pathway

    J. Biol. Chem.

    (2001)
  • A.M. Farrell et al.

    UVB irradiation up-regulates serine palmitoyltransferase in cultured human keratinocytes

    J. Lipid Res.

    (1998)
  • R.A. Memon et al.

    Regulation of sphingolipid and glycosphingolipid metabolism in extrahepatic tissues by endotoxin

    J. Lipid Res.

    (2001)
  • J.Y. Lehtonen et al.

    Activation of the de novo biosynthesis of sphingolipids mediates angiotensin II type 2 receptor-induced apoptosis

    J. Biol. Chem.

    (1999)
  • B. Liu et al.

    Glutathione regulation of neutral sphingomyelinase in tumor necrosis factor-alpha-induced cell death

    J. Biol. Chem.

    (1998)
  • K. Hidari et al.

    Complete removal of sphingolipids from the plasma membrane disrupts cell to substratum adhesion of mouse melanoma cells

    J. Biol. Chem.

    (1996)
  • G.P. Amarante-Mendes et al.

    Bcr-Abl exerts its antiapoptotic effect against diverse apoptotic stimuli through blockage of mitochondrial release of cytochrome C and activation of caspase-3

    Blood

    (1998)
  • E.C. Mandon et al.

    Subcellular localization and membrane topology of serine palmitoyltransferase, 3-dehydrosphinganine reductase, and sphinganine N-acyltransferase in mouse liver

    J. Biol. Chem.

    (1992)
  • Cited by (39)

    • Regulation of de novo sphingolipid biosynthesis by the ORMDL proteins and sphingosine kinase-1

      2015, Advances in Biological Regulation
      Citation Excerpt :

      Considerable evidence indicates that the regulation of de novo sphingolipid biosynthesis has important consequences for cellular physiology. For example biosynthesis of ceramide as a pro-apoptotic signal has been observed when cells are stressed with heat stress (Jenkins et al., 2002), sepsis and inflammatory cytokines (Memon et al., 1998), UV irradiation (Mullen et al., 2011; Panjarian et al., 2008; Uchida et al., 2003; Wispriyono et al., 2002), oxidative damage (Son et al., 2007) and the treatment of cells with a variety of chemotherapeutic compounds including daunorubicin (Bose et al., 1995), kinase and histone deacetylase inhibitors (Park et al., 2010), camptothecin and doxorubicin (Rath et al., 2009), cyclooxygenase inhibitors (Schiffmann et al., 2009), arsenic trioxide (Dbaibo et al., 2007), androgen ablation (Eto et al., 2003), fenretinide (Wang et al., 2001), and taxol (Charles et al., 2001; reviewed in Perry, 2000). Here we explore the mechanisms that control the biosynthesis of sphingolipids, particularly ceramide, to characterize the role of the ORMDLs and sphingosine kinase in the regulation of sphingolipid biosynthesis in both unstressed and stressed cells.

    View all citing articles on Scopus
    2

    These authors contributed equally to this article.

    1

    To whom reprint requests should be addressed at Department of Occupational and Environmental Health Sciences, Rm. 5142, Eugene Applebaum College of Pharmacy and Health Sciences, 259 Mack Ave., Detroit, MI 48201. Fax: (313) 577-0097. E-mail: [email protected].

    View full text