Skip to main content
SpringerLink
Log in

The inhibitory effect of dibutyryl cyclic AMP on docosahexaenoic acid-induced apoptosis in HL-60 cells through activation of the phosphatidylinositol-3 kinase pathway

  • Original Article
  • Published:
Environmental Health and Preventive Medicine Aims and scope

Abstract

Objective

Docosahexaenoic acid (DHA) is known as a chemopreventive substance for cancers. Previously we reported that DHA induces apoptosis in HL-60 cells. The aim of this study was to clarify the role of phosphatidylinositol 3-kinase (PI3-kinase)/Akt signaling during DHA-induced apoptosis in HL-60 cells.

Methods

The inhibitory effects of dibutyryl cAMP (db-cAMP) or LY294002 (a specific inhibitor of the PI3-kinase/Akt pathway) on DHA-induced apoptosis in HL-60 cells were evaluated by the appearance of apoptosis, and from the activities of caspases (3 and 8), the phospholylation of Akt, and cleavage of Bid using DNA indexes, emzymatic measurement of fragmented substrates, and Western blotting, respectively.

Results

The pre-incubation of db-cAMP reduced the activation of caspasses (3 and 8) during the occurrence of DHA-induced apoptosis in HL-60. However, the inhibition of PI3-kinase/Akt signaling by LY294002 resulted in recovery of the caspases’ activities, appearance of apoptotic cells, and cleavage of the Bid molecule when LY294002 was co-treated with db-cAMP before the occurrence of DHA-induced apoptosis in HL-60. It was also confirmed that LY294002 strongly inhibited phospholylation of Akt during db-cAMP induced-reduction of DHA-induced apoptosis in HL-60.

Conclusion

We demonstrated that DHA-induced apoptosis was sensitive to the modulation of PI3-kinase activity by treatment with db-cAMP or LY294002. These results may provide new insights into the mechanisms of the anti-cancer activity of DHA.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Rao VC, Hirose Y, Indranie C, Reddy SB. Modulation of experimental colon tumorigenesis by types and amounts of dietary fatty acids. Cancer Res. 2001; 61: 1927–1933.

    PubMed  CAS  Google Scholar 

  2. Latham P, Lund KE, Johnson TI. Dietary n-3 PUFA increases the apoptotic response to 1,2-dimethylhydrazine, reduces mitosis and suppresses the induction of carcinogenesis in the rat colon. Carcinogenesis. 1999; 20: 645–650.

    Article  PubMed  CAS  Google Scholar 

  3. Akbar M, Kim H. Protective effects of docosahexaenoic acid in staurosporine-induced apoptosis: involvement of phosphatidylinositol-3 kinase pathway. J Neurochem. 2002; 82: 655–665.

    Article  PubMed  CAS  Google Scholar 

  4. Calviello G, Palozza P, Piccioni E, Maggiano N, Frattucci A, Franceschelli P, Bartoli MG. Dietary supplementation with eicosapentaenoic and docosahexaenoic acid inhibits growth of morris hepatocarcinoma 3924A in rats: effects on proliferation and apoptosis. Int J Cancer. 1998; 75: 699–705.

    Article  PubMed  CAS  Google Scholar 

  5. Scheid MP, Lauener RW, Duronio V. Role of phosphatidylinositol 3-OH-kinase activity in the inhibition of apoptosis in haemopoietic cells: phosphatidylinositol 3-OH-kinase inhibitors reveal a difference in signaling between interleukin-3 and granulocyte-macrophage colony stimulating factor. Biochem J. 1995; 312: 159–162.

    PubMed  Google Scholar 

  6. Minshall C, Arkins S, Freund GG, Kelley KW. Requirement for phosphatidylinositol 3′-kinase to protect hemopoietic progenitors against apoptosis depends upon the extracellular survival factor. J Immunol. 1996; 156: 939–947.

    PubMed  CAS  Google Scholar 

  7. Songyang Z, Baltimore D, Cantley LC, Kaplan DR, Franke TF. Interleukin 3-dependent survival by the Akt protein kinase. Proc Natl Acad Sci USA. 1997; 94: 11345–11350.

    Article  PubMed  CAS  Google Scholar 

  8. Cynthia RLW, Anwer MS. Cyclic adenosine monophosphate-mediated protection against bile acid-induced apoptosis in cultured rat hepatocytes. Hepatology. 1998; 27: 1324–1331.

    Article  Google Scholar 

  9. Scheid MP, Foltz IN, Young PR, Schrader JW, Duronio V. Ceramide and cyclic adenosine monophosphate (cAMP) induce cAMP response element binding protein phosphorylation via distinct signaling pathways while having opposite effects on myeloid cell survival. Blood. 1999; 93: 217–225.

    PubMed  CAS  Google Scholar 

  10. Li M, Wang X, Meintzer MK, Laessig T, Birnbaum MJ, Heidenreich KA. Cyclic AMP promotes neuronal survival by phosphorylation of glycogen synthase kinase 3β. Mol Cell Biol. 2000; 20: 9356–9363.

    Article  PubMed  CAS  Google Scholar 

  11. Yusta B, Boushey RP, Drucker DJ. The glucagons-like peptide-2 receptor mediates direct inhibition of cellular apoptosis via a cAMP-dependent protein kinase-independent pathway. J Biol Chem. 2000; 275: 35345–35352.

    Article  PubMed  CAS  Google Scholar 

  12. Kim SN, Ahn Y-H, Kim SG, Park SD, Cho-Chung YS, Hong SH. 8-C1-cAMP induces cell cycle-specific apoptosis in human cancer cells. Int J Cancer. 2001; 93: 33–41.

    Article  PubMed  CAS  Google Scholar 

  13. Xia Z, Dickens M, Raingeaud J, Davis RJ, Greenberg ME. Opposing effects of ERK and JNK-p38 MAP kinase on apoptosis. Science. 1995; 270: 1326–1331.

    Article  PubMed  CAS  Google Scholar 

  14. Arita K, Kobuchi H, Utsumi T, Takehara Y, Akiyama J, Horton AA, Utsumi K. Mechanism of apootosis in HL-60 cells induced by n-3 and n-6 polyunsaturated fatty acids. Biochem Pharmacol. 2001; 62: 821–828.

    Article  PubMed  CAS  Google Scholar 

  15. Miura Y, Takahara K, Murata Y, Utsumi K, Tada T, Takahata K. Docosahexaenoic acid induces apoptosis via the Bax-independent pathway in HL-60 cells. Biosci Biotechnol Biochem. 2004; 68: 2415–2417.

    Article  PubMed  CAS  Google Scholar 

  16. Zang L, Himi T, Morita I, Murota S. Inhibition of phosphatidylinositol-3 kinase/Akt or mitogen-activated protein kinase signaling sensitizes endothelial cells to TNF-alpha cytotoxicity. Cell Death Differ. 2001; 8: 528–536.

    Article  Google Scholar 

  17. Kluck RM, Esposti MD, Perkins G, Renken C, Kuwana T, Bossy-Wetzel E, Goldberg M, Allen T, Barber MJ, Green DR, Newmeyer DD. The pro-apoptotic proteins. Bid and Bax. cause a limited permeabilization of the mitochondrial outer membrane that is enhanced by cytosol. J Cell Biol. 1999; 4: 809–822.

    Article  Google Scholar 

  18. Strokin M, Sergeeva M, Reiser G. Docosahezaenoic acid and arachidonic acid release in rat brain astrocytes is mediated by two separate isoforms of phospholipase A2 and is differently regulated by cyclic AMP and Ca2+. Br J Pharmacol. 2003; 139: 1014–1022.

    Article  PubMed  CAS  Google Scholar 

  19. Engler MM, Engler MB, Pierson DM, Molteni LB, Molteni A. Effects of docosahexaenoic acid on vascular pathology and reactivity in hypertension. Exp Biol Med. (Maywood) 2003; 228: 299–307.

    CAS  Google Scholar 

  20. Datta SR, Dudek H, Tao X, Masters S, Fu H, Gotoh Y, Greenberg ME. Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell. 1997; 91: 231–241.

    Article  PubMed  CAS  Google Scholar 

  21. Pawan K, April IM, Peter JP. p38 MAPK mediates γ-irradiation-induced endothelial cell apoptosis, and vascular endothelial growth factor protects endothelial cells through the phosphoinositide 3-kinase-Akt-Bcl-2 pathway. J Biol Chem. 2004; 279: 43352–43360.

    Article  Google Scholar 

  22. Sawada M, Nakashima S, Banno Y, Yamakawa H, Takenaka K, Shinoda J, Nishimura Y, Sakai N, Nozawa Y. Influence of Bax or Bcl-2 overexpression on the ceramide-dependent apoptotic pathway in glioma cells. Oncogene. 2000; 19: 3508–3520.

    Article  PubMed  CAS  Google Scholar 

  23. Gallagher R, Collins S, Trujillo J, McCredie K, Ahearn M, Tsai S, Metzgar R, Aulakh G, Ting R, Ruscetti, F, Gallo R. Characterization of the continuous, differentiating mycloid cell line (HL-60) from a patient with acute promyelocytic leukemia. Blood. 1979; 54: 713–733.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Graduate School of National Science and Technology, Okayama University, Okayama, Japan

    Yoshie Miura, Yoshiyuki Murata & Mikiro Tada

  2. Institute of Medical Science, Kurashiki Medical Center, Kurashiki, Japan

    Kozo Utsumi

  3. Faculty of Agriculture, Okayama University, Okayama, Japan

    Kyoya Takahata

  4. Department of Hygiene, Kawasaki Medical School, 577 Matsushima, 701-0192, Kurashiki, Japan

    Yoshie Miura & Takemi Otsuki

Authors
  1. Yoshie Miura
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yoshiyuki Murata
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kozo Utsumi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kyoya Takahata
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mikiro Tada
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Takemi Otsuki
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yoshie Miura.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Miura, Y., Murata, Y., Utsumi, K. et al. The inhibitory effect of dibutyryl cyclic AMP on docosahexaenoic acid-induced apoptosis in HL-60 cells through activation of the phosphatidylinositol-3 kinase pathway. Environ Health Prev Med 10, 184–189 (2005). https://doi.org/10.1007/BF02897709

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02897709

Key words

Search

Navigation