Skip to main content
PMC home page
Immunology logoLink to Immunology
. 1997 Aug;91(4):594–600. doi: 10.1046/j.1365-2567.1997.d01-2281.x

An immunosuppressive agent, FTY720, increases intracellular concentration of calcium ion and induces apoptosis in HL-60.

T Shinomiya 1, X K Li 1, H Amemiya 1, S Suzuki 1
PMCID: PMC1363881  PMID: 9378500

Abstract

We previously reported that FTY720 is an efficient inducer of apoptosis in lymphocytes and cultured cell lines. In the present study, HL-60 human promyerocytoma cells also induced apoptosis through in vitro treatment with the drug, demonstrating extensive DNA fragmentation 6 hr after incubation. The major target of FTY720 was the common signalling pathway of apoptosis, since a rapid (< 1 min) increase in the intracellular Ca2+ concentration ([Ca2+]i) was found in the cells treated with the drug. Calcium chelation in the culture medium with EGTA did not affect the [Ca2+]i mobilization. A phospholipase C inhibitor, U73122, inhibited the increase in [Ca2+]i as well as the fragmentation of the nuclear DNA, whereas U73343, a non-effective analogue of U73122, had little effect. These results suggest that FTY720-induced apoptosis is mediated through an activation of phospholipase C and the subsequent release of Ca2+ from intracellular calcium pools. In addition, the treatment of HL-60 with pertussis toxin (PTX) did not inhibit Ca2+ mobilization or apoptosis, suggesting that the activation of phospholipase C is independent of PTX-sensitive G-proteins.

Full text

PDF
594

Images in this article

595Figure 1
on p.595
597Figure 6
on p.597

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alley M. C., Scudiero D. A., Monks A., Hursey M. L., Czerwinski M. J., Fine D. L., Abbott B. J., Mayo J. G., Shoemaker R. H., Boyd M. R. Feasibility of drug screening with panels of human tumor cell lines using a microculture tetrazolium assay. Cancer Res. 1988 Feb 1;48(3):589–601. [PubMed] [Google Scholar]
  2. Aragozzini F., Manachini P. L., Craveri R., Rindone B., Scolastico C. Structure of thermozymocidin. Experientia. 1972 Aug 15;28(8):881–882. doi: 10.1007/BF01924918. [DOI] [PubMed] [Google Scholar]
  3. Arends M. J., Morris R. G., Wyllie A. H. Apoptosis. The role of the endonuclease. Am J Pathol. 1990 Mar;136(3):593–608. [PMC free article] [PubMed] [Google Scholar]
  4. Berridge M. J. Inositol trisphosphate and calcium signalling. Nature. 1993 Jan 28;361(6410):315–325. doi: 10.1038/361315a0. [DOI] [PubMed] [Google Scholar]
  5. Berridge M. J. Inositol trisphosphate and diacylglycerol: two interacting second messengers. Annu Rev Biochem. 1987;56:159–193. doi: 10.1146/annurev.bi.56.070187.001111. [DOI] [PubMed] [Google Scholar]
  6. Bleasdale J. E., Thakur N. R., Gremban R. S., Bundy G. L., Fitzpatrick F. A., Smith R. J., Bunting S. Selective inhibition of receptor-coupled phospholipase C-dependent processes in human platelets and polymorphonuclear neutrophils. J Pharmacol Exp Ther. 1990 Nov;255(2):756–768. [PubMed] [Google Scholar]
  7. Collins S. J., Gallo R. C., Gallagher R. E. Continuous growth and differentiation of human myeloid leukaemic cells in suspension culture. Nature. 1977 Nov 24;270(5635):347–349. doi: 10.1038/270347a0. [DOI] [PubMed] [Google Scholar]
  8. Fujita T., Inoue K., Yamamoto S., Ikumoto T., Sasaki S., Toyama R., Chiba K., Hoshino Y., Okumoto T. Fungal metabolites. Part 11. A potent immunosuppressive activity found in Isaria sinclairii metabolite. J Antibiot (Tokyo) 1994 Feb;47(2):208–215. doi: 10.7164/antibiotics.47.208. [DOI] [PubMed] [Google Scholar]
  9. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  10. Hannun Y. A., Loomis C. R., Merrill A. H., Jr, Bell R. M. Sphingosine inhibition of protein kinase C activity and of phorbol dibutyrate binding in vitro and in human platelets. J Biol Chem. 1986 Sep 25;261(27):12604–12609. [PubMed] [Google Scholar]
  11. Harman A. W., Maxwell M. J. An evaluation of the role of calcium in cell injury. Annu Rev Pharmacol Toxicol. 1995;35:129–144. doi: 10.1146/annurev.pa.35.040195.001021. [DOI] [PubMed] [Google Scholar]
  12. Kaiser N., Edelman I. S. Calcium dependence of glucocorticoid-induced lymphocytolysis. Proc Natl Acad Sci U S A. 1977 Feb;74(2):638–642. doi: 10.1073/pnas.74.2.638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Katada T., Ui M. Direct modification of the membrane adenylate cyclase system by islet-activating protein due to ADP-ribosylation of a membrane protein. Proc Natl Acad Sci U S A. 1982 May;79(10):3129–3133. doi: 10.1073/pnas.79.10.3129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Katada T., Ui M. Islet-activating protein. Enhanced insulin secretion and cyclic AMP accumulation in pancreatic islets due to activation of native calcium ionophores. J Biol Chem. 1979 Jan 25;254(2):469–479. [PubMed] [Google Scholar]
  15. Kluepfel D., Bagli J., Baker H., Charest M. P., Kudelski A. Myriocin, a new antifungal antibiotic from Myriococcum albomyces. J Antibiot (Tokyo) 1972 Feb;25(2):109–115. doi: 10.7164/antibiotics.25.109. [DOI] [PubMed] [Google Scholar]
  16. Liu Y. J., Mason D. Y., Johnson G. D., Abbot S., Gregory C. D., Hardie D. L., Gordon J., MacLennan I. C. Germinal center cells express bcl-2 protein after activation by signals which prevent their entry into apoptosis. Eur J Immunol. 1991 Aug;21(8):1905–1910. doi: 10.1002/eji.1830210819. [DOI] [PubMed] [Google Scholar]
  17. Martel R. R., Klicius J., Galet S. Inhibition of the immune response by rapamycin, a new antifungal antibiotic. Can J Physiol Pharmacol. 1977 Feb;55(1):48–51. doi: 10.1139/y77-007. [DOI] [PubMed] [Google Scholar]
  18. McConkey D. J., Nicotera P., Hartzell P., Bellomo G., Wyllie A. H., Orrenius S. Glucocorticoids activate a suicide process in thymocytes through an elevation of cytosolic Ca2+ concentration. Arch Biochem Biophys. 1989 Feb 15;269(1):365–370. doi: 10.1016/0003-9861(89)90119-7. [DOI] [PubMed] [Google Scholar]
  19. Miyake Y., Kozutsumi Y., Nakamura S., Fujita T., Kawasaki T. Serine palmitoyltransferase is the primary target of a sphingosine-like immunosuppressant, ISP-1/myriocin. Biochem Biophys Res Commun. 1995 Jun 15;211(2):396–403. doi: 10.1006/bbrc.1995.1827. [DOI] [PubMed] [Google Scholar]
  20. Nicotera P., Bellomo G., Orrenius S. Calcium-mediated mechanisms in chemically induced cell death. Annu Rev Pharmacol Toxicol. 1992;32:449–470. doi: 10.1146/annurev.pa.32.040192.002313. [DOI] [PubMed] [Google Scholar]
  21. Ochiai T., Nakajima K., Nagata M., Hori S., Asano T., Isono K. Studies of the induction and maintenance of long-term graft acceptance by treatment with FK506 in heterotopic cardiac allotransplantation in rats. Transplantation. 1987 Dec;44(6):734–738. doi: 10.1097/00007890-198712000-00002. [DOI] [PubMed] [Google Scholar]
  22. Ohta H., Sweeney E. A., Masamune A., Yatomi Y., Hakomori S., Igarashi Y. Induction of apoptosis by sphingosine in human leukemic HL-60 cells: a possible endogenous modulator of apoptotic DNA fragmentation occurring during phorbol ester-induced differentiation. Cancer Res. 1995 Feb 1;55(3):691–697. [PubMed] [Google Scholar]
  23. Ohta H., Sweeney E. A., Masamune A., Yatomi Y., Hakomori S., Igarashi Y. Induction of apoptosis by sphingosine in human leukemic HL-60 cells: a possible endogenous modulator of apoptotic DNA fragmentation occurring during phorbol ester-induced differentiation. Cancer Res. 1995 Feb 1;55(3):691–697. [PubMed] [Google Scholar]
  24. Ohta H., Yatomi Y., Sweeney E. A., Hakomori S., Igarashi Y. A possible role of sphingosine in induction of apoptosis by tumor necrosis factor-alpha in human neutrophils. FEBS Lett. 1994 Dec 5;355(3):267–270. doi: 10.1016/0014-5793(94)01218-0. [DOI] [PubMed] [Google Scholar]
  25. Okajima F., Tomura H., Sho K., Nochi H., Tamoto K., Kondo Y. Involvement of pertussis toxin-sensitive GTP-binding proteins in sphingosine 1-phosphate-induced activation of phospholipase C-Ca2+ system in HL60 leukemia cells. FEBS Lett. 1996 Feb 5;379(3):260–264. doi: 10.1016/0014-5793(95)01526-4. [DOI] [PubMed] [Google Scholar]
  26. Perotti M., Toddei F., Mirabelli F., Vairetti M., Bellomo G., McConkey D. J., Orrenius S. Calcium-dependent DNA fragmentation in human synovial cells exposed to cold shock. FEBS Lett. 1990 Jan 1;259(2):331–334. doi: 10.1016/0014-5793(90)80040-p. [DOI] [PubMed] [Google Scholar]
  27. Quesniaux V. F., Schreier M. H., Wenger R. M., Hiestand P. C., Harding M. W., Van Regenmortel M. H. Cyclophilin binds to the region of cyclosporine involved in its immunosuppressive activity. Eur J Immunol. 1987 Sep;17(9):1359–1365. doi: 10.1002/eji.1830170921. [DOI] [PubMed] [Google Scholar]
  28. Rüegger A., Kuhn M., Lichti H., Loosli H. R., Huguenin R., Quiquerez C., von Wartburg A. Cyclosporin A, ein immunsuppressiv wirksamer Peptidmetabolit aus Trichoderma polysporum (Link ex Pers.) Rifai. Helv Chim Acta. 1976;59(4):1075–1092. doi: 10.1002/hlca.19760590412. [DOI] [PubMed] [Google Scholar]
  29. Sasaguri T., Hirata M., Itoh T., Koga T., Kuriyama H. Guanine nucleotide binding protein involved in muscarinic responses in the pig coronary artery is insensitive to islet-activating protein. Biochem J. 1986 Nov 1;239(3):567–574. doi: 10.1042/bj2390567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Suzuki S., Enosawa S., Kakefuda T., Shinomiya T., Amari M., Naoe S., Hoshino Y., Chiba K. A novel immunosuppressant, FTY720, with a unique mechanism of action, induces long-term graft acceptance in rat and dog allotransplantation. Transplantation. 1996 Jan 27;61(2):200–205. doi: 10.1097/00007890-199601270-00006. [DOI] [PubMed] [Google Scholar]
  31. Suzuki S., Li X. K., Enosawa S., Shinomiya T. A new immunosuppressant, FTY720, induces bcl-2-associated apoptotic cell death in human lymphocytes. Immunology. 1996 Dec;89(4):518–523. doi: 10.1046/j.1365-2567.1996.d01-777.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Suzuki S., Li X. K., Shinomiya T., Enosawa S., Amemiya H., Amari M., Naoe S. The in vivo induction of lymphocyte apoptosis in MRL-lpr/lpr mice treated with FTY720. Clin Exp Immunol. 1997 Jan;107(1):103–111. doi: 10.1046/j.1365-2249.1997.d01-885.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tanaka H., Kuroda A., Marusawa H., Hashimoto M., Hatanaka H., Kino T., Goto T., Okuhara M. Physicochemical properties of FK-506, a novel immunosuppressant isolated from Streptomyces tsukubaensis. Transplant Proc. 1987 Oct;19(5 Suppl 6):11–16. [PubMed] [Google Scholar]
  34. Thastrup O., Cullen P. J., Drøbak B. K., Hanley M. R., Dawson A. P. Thapsigargin, a tumor promoter, discharges intracellular Ca2+ stores by specific inhibition of the endoplasmic reticulum Ca2(+)-ATPase. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2466–2470. doi: 10.1073/pnas.87.7.2466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Thastrup O., Foder B., Scharff O. The calcium mobilizing tumor promoting agent, thapsigargin elevates the platelet cytoplasmic free calcium concentration to a higher steady state level. A possible mechanism of action for the tumor promotion. Biochem Biophys Res Commun. 1987 Feb 13;142(3):654–660. doi: 10.1016/0006-291x(87)91464-1. [DOI] [PubMed] [Google Scholar]
  36. Torigoe T., Millan J. A., Takayama S., Taichman R., Miyashita T., Reed J. C. Bcl-2 inhibits T-cell-mediated cytolysis of a leukemia cell line. Cancer Res. 1994 Sep 15;54(18):4851–4854. [PubMed] [Google Scholar]
  37. Vézina C., Kudelski A., Sehgal S. N. Rapamycin (AY-22,989), a new antifungal antibiotic. I. Taxonomy of the producing streptomycete and isolation of the active principle. J Antibiot (Tokyo) 1975 Oct;28(10):721–726. doi: 10.7164/antibiotics.28.721. [DOI] [PubMed] [Google Scholar]
  38. Wyllie A. H. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature. 1980 Apr 10;284(5756):555–556. doi: 10.1038/284555a0. [DOI] [PubMed] [Google Scholar]

Articles from Immunology are provided here courtesy of British Society for Immunology

RESOURCES