Apoptosis

, Volume 4, Issue 4, pp 255–261

Fas-mediated apoptosis in Jurkat cells is suppressed in the pre-G2/M phase

  • N. Hiroi
  • H. Maruta
  • S. Tanuma
Article

Abstract

The relationship between the cell cycle and Fas-mediated apoptosis was investigated using Jurkat cells. Analysis of the inducibility of apoptosis by anti-Fas antibody during the cell cycle synchronized by the thymidine double-block method, showed that apoptosis was induced in only 50% of the G2/M phase cells, while most of cells in the other phases underwent apoptosis. These observations indicate that G2/M phase cells are more resistant to Fas-mediated apoptosis than cells in other phases. Furthermore, a detailed analysis of G2/M phase found that only 20–30% of the cells underwent apoptosis 12 h after the removal of the second thymidine block (pre-G2/M phase). This suggests that Fas-mediated apoptosis is potently suppressed during the pre-G2/M phase. A possible explanation for the observation that cells in the pre-G2/M phase are less sensitive to anti-Fas antibody is lower expression level of Fas. To test this possibility, Fas expression levels on the cell surface during the cell cycle were examined. The content of Fas on the cell surface, however, did not change appreciably during the cell cycle. Thus, the suppression of apoptosis in the pre-G2/M phase is determined downstream after the receipt of the apoptotic signal through Fas.

Apoptosis cell cycle G2/M phase Fas Jurkat cell. 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Wyllie AH, Kerr JFR, Currie AR. Cell death: the significance of apoptosis. Int Rev Cytol 1980; 68: 251-306.Google Scholar
  2. 2.
    King KL, Cidlowski JA. Cell cycle and apoptosis: common pathways to life and death. J Cell Biochem 1995; 58: 175-180.Google Scholar
  3. 3.
    Nagata S. Apoptosis by death factor. Cell 1997; 88: 355-365.Google Scholar
  4. 4.
    Miyawaki T, Uehara T, Tsuji T, et al. Differential expression of apoptosis-related CD95 antigen on lymphocyte subpopulations in human peripheral blood. J Immunol 1992; 149: 3753-3760.Google Scholar
  5. 5.
    Klas C, Debatin KM, Jonker R, Krammer PH. Activation interferes with the APO-1 pathway in mature human T cells. Int Immunol 1993; 5: 625-630.Google Scholar
  6. 6.
    Beletskaya IV, Nikonova LV, Beletsky IP. Cell cycle specificity of Fas-mediated apoptosis in WIL-2 cells. FEBS Lett 1997; 412: 91-93.Google Scholar
  7. 7.
    Dao T, Huleatt JW, Hingorani R, Crispe N. Specific resistance of T cells to CD95-induced apoptosis during S phase of the cell cycle. J Immunol 1997; 159: 4261-4267.Google Scholar
  8. 8.
    Li-Weber M, Salgame P, Krammer PH, et al. Differential interaction of nuclear factors with the PRE-1 enhancer element of human IL-4 promoter in different T cell subsets. Blood 1997; 97: 1194-1200.Google Scholar
  9. 9.
    Juo P, Kuo CJ, Reynols SE, et al. Fas activation of the p38 mitogen-activated protein kinase signaling pathway requires ICE/CED-3 family proteases. Mol Cell Biol 1997; 17: 24-35.Google Scholar
  10. 10.
    Lenczowski JM, Dominguez L, Eder AM, et al. Lack of a role for Jun kinase and AP-1 in Fas-induced apoptosis. Mol Cell Biol 1997; 17: 170-181.Google Scholar
  11. 11.
    Ponton A, Clément M-V, Stamenkovic I. The CD95 (APO-1/Fas) receptor activates NF-kB independently of its cytotoxic function. J Biol Chem 1996; 271: 8991-8995.Google Scholar
  12. 12.
    Eischen CM, Dick CJ, Leibson J. Tyrosine kinase activation provides an early and requisite signal for Fas-induced apoptosis. J Immunol 1994; 153: 1947-1954.Google Scholar
  13. 13.
    Shimizu T, O'Connor PM, Kohn KW, Pommier Y. Unscheduled activation of Cyclin B1/Cdc2 kinase in human promyelocytic leukemia cell line HL60 cells undergoing apoptosis induced by DNA damage. Cancer Res 1995; 55: 228-231.Google Scholar
  14. 14.
    Shiokawa D, Ohyama H, Yamada T, Takahashi K, Tanuma S. Identification of an endonuclease responsible for apoptosis in rat thymocytes. Eur J Biochem 1994; 226: 23-30.Google Scholar
  15. 15.
    Tanuma S, Shiokawa D. Multiple forms of nuclear deoxyribonuclease in rat thymocytes. Biochem Biophys Res Comm 1994; 203: 789-797.Google Scholar
  16. 16.
    Huschtscha LI, Jeitner TM, Andersson CE, Bartier WA, Tattersall MHN. Identification of apoptotic and necrotic human leukemic cells by flow cytometry. Exp Cell Res 1994; 212: 161-165.Google Scholar
  17. 17.
    Lacombe F, Belloc F. Flow cytometry study of cell cycle, apoptosis and drug resistance in acute leukemia. Hematol Cell Ther 1996; 38: 495-504.Google Scholar
  18. 18.
    Cheng J, Zhou T, Liu C, et al. Protection from Fas-mediated apoptosis by a soluble from of the Fas molecule. Science 1994; 263: 1759-1762.Google Scholar
  19. 19.
    Schattner EJ, Elkon KB, Yoo D, et al. CD40 ligation induces apo-1/Fas expression on human B lymphocytes and facilitates apoptosis through the Apo-1/Fas pathway. J Exp Med 1995; 182: 1557-1565.Google Scholar
  20. 20.
    Rathmell JC, Townsend SE, Xu JC, et al. Expansion or elimination of B cells in vivo: dual roles for CD40-and Fas (CD95)-ligands modulated by the B cell antigen receptor. Cell 1996; 87: 319-329.Google Scholar
  21. 21.
    Watanabe-Fukunaga R, Brannan CI, Copeland NG, Jenkins NA, Nagata S. Lymphoproliferation disorder in mice explained by defects in Fas antigen that mediates apoptosis. Nature 1992; 356: 314-317.Google Scholar
  22. 22.
    Takahashi, T, Tanaka M, Brannan CI, et al. Generalized lymphoproliferative disease in mice, caused by a point mutation in the Fas ligand. Cell 1994; 76: 969-976.Google Scholar
  23. 23.
    Alderson MR, Armitage RJ, Maraskovsky E, et al. Fas transduces activation signals in normal human T lymphocytes. J Exp Med. 1993; 178: 2231-2235.Google Scholar
  24. 24.
    Dianzani U, Bragardo M, Ramenghi U, et al. Deficiency of the Fas apoptosis pathway without Fas gene mutations in pediatric patients with autoimmunity/lymphoproliferation. Blood 1997; 89: 2871-2879.Google Scholar
  25. 25.
    Puck JM, Sneller MC. ALPS: an autoimmune human lymphoproliferative syndrome associated with abnormal lymphocyte apoptosis. Semin Immunol 1997; 9: 77-84.Google Scholar
  26. 26.
    Li F, Ambrosini G, Altieri DC, et al. Control of apoptosis and mitotic spindle checkpoint by survivin. Nature 1998; 396: 580-584.Google Scholar
  27. 27.
    Furukawa Y, Iwase A, Terui J, et al. Transcriptional activation of the cdc2 gene is associated with Fas-induced apoptosis of human hematopoietic cells. J Biol Chem 1996; 271: 28469-28477.Google Scholar
  28. 28.
    Kitagawa M, Okabe T, Yasuda H, et al. Butyrolactone I, a selective inhibitor of cdk2 and cdc2 kinase. Oncogene 1993; 8: 2425-2432.Google Scholar
  29. 29.
    Salvesen GS, Dixit VM. Caspases: intercellular signaling by proteolysis. Cell 1997; 91: 443-446.Google Scholar
  30. 30.
    Shiokawa D, Nishimura K, Maruta H, Tanuma S. DNA fragmentation during thymic apoptosis is catalyzed by DNase γ. Apoptosis 1996; 1: 147-152.Google Scholar
  31. 31.
    Enari M, Sakahira H, Nagata S, et al. A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 1998; 391: 43-50.Google Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • N. Hiroi
    • 1
  • H. Maruta
    • 1
  • S. Tanuma
    • 1
  1. 1.Department of Biochemistry, Faculty of Pharmaceutical SciencesScience University of TokyoShinjuku-ku, TokyoJapan

Personalised recommendations