• programmed cell death;
  • biomembrane;
  • cholesterol;
  • lipoxygenase;
  • phospholipase A2


  1. Top of page
  2. Abstract
  3. References

Transforming growth factor β1 (TGFβ1) and cisplatin induce apoptosis (programmed cell death, PCD) in human erythroleukemia K562 cells in an additive manner. After PCD was induced in K562 cells, analysis of phospholipid composition, fatty acids and cholesterol content in their membranes showed a decrease in phosphatidylethanolamine and an increase in phosphatidylserine, cardiolipin and phosphatidic acid. Moreover, cisplatin but not TGFβ1 enhanced sphingomyeline levels in apoptotic cells, whereas TGFβ1 increased the amount of linoleic acid and, more remarkably, of cholesterol. The combination TGFβ+ cisplatin produced membrane changes similar to those provoked by each inducer individually. Furthermore, the specific activities of 5-lipoxygenase and cytosolic phospholipase A2, both modulating the physical properties of membranes and membrane-lipid-mediated intracellular signalling, were enhanced by treatment with TGFβ1 or TGFβ1 + cisplatin. These findings highlight the profound changes in cell membranes during the biochemical events of the apoptotic pathway.


programmed cell death


transforming growth factor β1


secretory phospholipase A2


cytosolic phospholipase A2


arachidonic acid


5-hydroperoxyarachidonic acid


gas chromatography-mass spectrometry


Lipoxygenase (EC


phospholipase A2 (EC


  1. Top of page
  2. Abstract
  3. References
  • 1
    Liepins, A. & Bustamante, J. O. (1994) Cell injury and apoptosis, Scan. Microscopy 8, 631643.
  • 2
    Hale, A. J., Smith, C. A., Sutherland, L. C., Stoneman, V. E. A., Longthorne, V. L., Culhane, A. C. & Williams, G. T. (1996) Apoptosis: molecular regulation of cell death, Eur. J. Biochem. 236, 126.
  • 3
    Sweeley, C. C. (1994) Lipids and the modulation of cell function, Curr. Topics Membr. 40, 357360.
  • 4
    El-Mofty, S. K., Scrutton, M. C., Serroni, A., Nicolini, C. & Farber, J. L. (1975) Early, reversible plasma membrane injury in galactosamine-induced liver cell death, Am. J. Pathol 79, 579595.
  • 5
    Savill, J. S., Wyllie, A. H., Henson, J. E., Walport, M. J., Henson, P. M. & Haslett, C. (1989) Macrophage phagocytosis of aging neutrophils in inflammation. Programmed cell death in the neutro-phil leads to its recognition by macrophages, J. Clin. Invest. 83, 865875.
  • 6
    Motyl, T., Kasterka, M., Grzelkowska, K., Blachowski, S. & Sysa, P. (1993) TGF-β1 inhibits polyamine biosynthesis in K562 leukemic cells, Ann. Hematol. 67, 285288.
  • 7
    Marushige, K. & Marushige, Y. (1994) Induction of apoptosis by transforming growth factor β1 in glioma and trigeininal neurinoma cells, Anticancer Res 14, 24192424.
  • 8
    Ewen, M. E., Sluss, H. K., Whitehouse, L. L. & Livingstone, D. M. (1993) TGFβ inhibition of Cdk4 synthesis is linked to cell cycle arrest, Cell 74, 10091020.
  • 9
    Piacentini, M., Fesus, L. & Melino, G. (1993) Multiple cell cycle access to the apoptotic death programme in human neuroblastoma cells, FEBS Lett. 320, 150154.
  • 10
    Navakova, O., Vrana, O., Kiseleva, V. I. & Brabec, V. (1995) DNA interactions of antitumor platinum(IV) complexes, Eur. J. Biochem. 228, 616624.
  • 11
    Kuhn, H., Belkner, J., Wiesner, R. & Brash, A. R. (1990) Oxygenation of biological membranes by the pure reticulocyte lipoxygenase, J. Biol. Chem. 265, 1835118361.
  • 12
    Regdel, D., Schewe, T. & Kuehn, H. (1994) Tomato fruit lipoxygenase: isolation, partial purification, characterization, and interaction with biological membranes, Biochemistry (Moscow) 59, 581586.
  • 13
    Maccarrone, M., Van Aarle, P. G. M., Veldink, G. A. & Vliegenthart, J. F. G. (1994) In vitro oxygenation of soybean biomembranes by lipoxygenase-2, Biochim. Biophys. Acta 1190, 164169.
  • 14
    Los, M., Schenk, H., Hexel, K., Baeuerle, P. A., Droge, W. & Schulze-Osthoff, K. (1995) IL-2 gene expression and NF-kB activation through CD28 requires reactive oxygen production by 5-lipoxygenase, EMBO J. 14, 37313740.
  • 15
    Schalkwijk, C. G., De Vet, E., Pfeilschiftcr, J. & Van den Bosch, H. (1992) Interleukin-1β and transforming growth factor-β2 enhance cytosolic high-molecular mass phospholipase A2 activity and induce prostaglandin E2 formation in rat mesangial cells, Eur. J. Biochem. 210, 169176.
  • 16
    Schalkwijk, C. G., Spaargaren, M., Defize, L. H. K., Verkleij, A. J., Van den Bosch, H. & Boonstra, J. (1995) Epidermal growth factor (EGF) induces serine phosphorylation-dependent activation and calcium-dependent translocation of the cytosolic phospholipase A2, Eur. J. Biochem. 231, 593601.
  • 17
    Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal. Biochem. 72, 248254.
  • 18
    Leist, M., Gartner, F., Bohlinger, I., Tiegs, G. & Wendel, A. (1994) Application of the cell death detection ELISA for the detection of tumor necrosis factor-induced DNA fragmentation in murine models of inflammatory organ failure, Biochemica 11, 2022.
  • 19
    Brungs, M., Radmark, O., Samuelsson, B. & Steinhilber, D. (1995) Sequential induction of 5-lipoxygenase gene expression and activity in Mono Mac 6 cells by transforming growth factor β and 1,25-dihydroxyvitamin D3, Proc. Natl Acad. Sci. USA 92, 107111.
  • 20
    Vervoordeldonk, M. J. B. M., Schalkwijk, C. G., Vishwanath, B. S., Aarsman, A. J. & Van den Bosch, H. (1994) Levels and localization of group II phospholipase A2 and annexin I in interleukin-and dexamethasone-treated rat mesangial cells: evidence against annexin mediation of the dexamethasone-induced inhibition of group II phospholipases A2, Biochem. Biophys. Acta 1224, 541550.
  • 21
    Hauser, H. & Lipka, G. (1994) Lipid dynamics in brush border membrane, Curr. Topics Membr. 40, 167195.
  • 22
    Fadok, V. A., Voelker, D. R., Campbell, P. A., Cohen, J. J., Bratton, D. L. & Henson, P. M. (1992) Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages, J. Immunol. 148, 22072216.
  • 23
    Sambrano, G. R. & Steinberg, D. (1995) Recognition of oxidatively damaged and apoptotic cells by an oxidized low density lipoprotein receptor on mouse peritoneal macrophages: role of membrane phosphatidylserine, Proc. Natl Acad. Sci. USA 92, 13961400.
  • 24
    Briehl, M. M., Cotgreave, I. A. & Powis, G. (1995) Downregulation of the antioxidant defence during glucocorticoid-mediated apoptosis, Cell Death Differ 2, 4146.
  • 25
    Verheij, M., Bose, R., Lin, X.-H., Yao, B., Jarvis, W. D., Grant, S., Birrer, M. J., Szabo, E., Zon, L. I., Kyriakis, J. M., Haimovitz-Friedman, A., Fuks, Z. & Kolesnick, R. N. (1996) Requirement for ceramide-initiated SAPK/JNK signalling in stress-induced apoptosis, Nature 380, 7579.
  • 26
    Slotte, J. P., Porn, M. I. & Harmala, A.-S. (1994) Flow and distribution of cholesterol – Effects of phospholipids, Curr. Topics Membr. 40, 483502.
  • 27
    Parasassi, T., Di Stefmo, M., Loiero, M., Ravagnan, G. & Gratton, E. (1994) Cholesterol modifies water concentration and dynamics in phospholipid bilayers. A fluorescence study using Laurdan probe, Biophys. J. 66, 763766.
  • 28
    Casey, P. J. (1995) Protein lipidation in cell signaling, Science 268, 221225.
  • 29
    Sandstrom, P. A., Tebbey, P. W., Van Cleave, S. & Buttke, T. M. (1994) Lipid hydroperoxides induce apoptosis in T cells displaying a HIV-associated glutathione peroxidase deficiency, J. Biol. Chem. 269, 798801.
  • 30
    Liscovitch, M. & Cantley, L. C. (1994) Lipid second messengers, Cell 77, 329334.