Abstract
The IPC-81 myeloid leukaemia cells undergo apoptosis rapidly after cAMP stimulation (6 h) and cell death is prevented by early over-expression of the cAMP-inducible transcription repressor ICER, that blocks cAMP-dependent nuclear signalling. Therefore, the expression of specific genes controlled by CRE-containing promoters is likely to determine cell fate. We now show that cAMP-induced cell death also is abrogated by the over-expression of the anti-apoptotic gene, Bcl-2. Contrary to ICER, Bcl-2 does not affect cAMP-signalling and allows the analysis of cAMP responses in death rescued cells. The Bcl-2 transfected cells treated with 8-CPT-cAMP were growth-arrested and thereafter cells embarked in granulocytic differentiation, with no additional stimulation. Neutrophilic polynuclear granulocytes benefited from a long life span in G0-G1 and remained functional (phagocytosis). This work demonstrates that, using anti-apoptosis regulators, ‘death signals’ could be exploited to trigger distinct biological responses. Indeed, cAMP signal can trigger several simultaneously developing biological programs, in the same cell, i.e., growth regulation, apoptosis and differentiation. This cell system should prove useful to determine how a tumour cell can be re-programmed for either apoptosis or functional maturation by physiological signals. Cell Death and Differentiation (2000) 7, 1081–1089.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- cAMP:
-
cyclic adenosine monophosphate
- 8-CPT-cAMP:
-
8-chloro-phenyl-thio-cAMP
- CRE:
-
c-AMP responsive element
- FACS:
-
fluorescence-activated cell sorting
- PKC:
-
Protein kinase C
- PKA:
-
c-AMP-dependent protein kinase
- RAR:
-
retinoic acid receptor
- RXR:
-
retinoid-X-receptor
- TUNEL:
-
terminal deoxynucleotide transferase-mediated dUTP-biotin nick-end labelling
References
Arends MJ and Wyllie AH . 1991 Apoptosis: mechanisms and roles in pathology. Int. Rev. Exp. Pathol. 32: 223–254
Fisher DE . 1994 Apoptosis in cancer therapy: crossing the threshold. Cell 78: 539–542
Hoffman B and Liebermann DA . 1994 Molecular controls of apoptosis: differentiation/growth arrest primary response genes, proto-oncogenes, and tumor suppressor genes as positive and negative modulators. Oncogene 9: 1807–1812
White E . 1996 Life, death, and the pursuit of apoptosis. Genes Dev. 10: 1–15
Rowan S and Fisher DE . 1997 Mechanisms of apoptotic cell death. Leukemia 11: 457–465
Savill JS, Wyllie AH, Henson JE, Walport MJ, Henson PM and Haslett C . 1989 Macrophage phagocytosis of aging neutrophils in inflammation. Programmed cell death in the neutrophil leads to its recognition by macrophages. J. Clin. Invest. 83: 865–875
Zamzami N, Brenner C, Marzo I, Susin SA and Kroemer G . 1998 Subcellular and submitochondrial mode of action of Bcl-2-like oncoproteins. Oncogene 16: 2265–2282
Reed J . 1998 Bcl-2 family proteins. Oncogene 17: 3225–3236
Chao DT and Korsmeyer SJ . 1998 Bcl-2 family: Regulators of Cell Death. Annu. Rev. Immunol. 16: 395–419
Hockenbery DM . 1994 Bcl-2 in cancer, development and apoptosis. J. Cell. Sci. (Suppl.) 18: 51–55
Cory S, Harris AW and Strasser A . 1994 Insights from transgenic mice regarding the role of bcl-2 in normal and neoplastic lymphoid cells. Philos. Trans. R. Soc. Lond. B. Biol. Sci. 345: 289–295
Merry DE and Korsmeyer SJ . 1997 Bcl-2 gene family in the nervous system. Annu. Rev. Neurosci. 20: 245–267
Hawkins CJ and Vaux DL . 1997 The role of the Bcl-2 family of apoptosis regulatory proteins in the immune system. Semin. Immunol. 9: 25–33
Dragovich T, Rudin CM and Thompson CB . 1998 Signal transduction pathways that regulate cell survival and cell death. Oncogene 17: 3207–3213
Naumovski L and Cleary ML . 1994 Bcl2 inhibits apoptosis associated with terminal differentiation of HL-60 myeloid leukemia cells. Blood 83: 2261–2267
Bruel A, Benoit G, De Nay D, Brown S and Lanotte M . 1995 Distinct apoptotic responses in maturation sensitive and resistant t(15;17) acute promyelocytic leukemia NB4 cells. 9-cis retinoic acid induces apoptosis independent of maturation and Bcl-2 expression. Leukemia 9: 1173–1184
Nagy L, Thomazy VA, Shipley GL, Fesus L, Lamph W, Heyman RA, Chandraratna RA and Davies PJ . 1995 Activation of retinoid X receptors induces apoptosis in HL-60 cell lines. Mol. Cell. Biol. 15: 3540–3551
Grillier I, Umiel T, Elstner E, Collins SJ and Koeffler HP . 1997 Alterations of differentiation, clonal proliferation, cell cycle progression and bcl-2 expression in RAR alpha-altered sublines of HL-60. Leukemia 11: 393–400
O'Reilly LA, Huang DC and Strasser A . 1996 The cell death inhibitor Bcl-2 and its homologues influence control of cell cycle entry. EMBO J. 15: 6979–6990
Huang DC, O'Reilly LA, Strasser A and Cory S . 1997 The anti-apoptosis function of Bcl-2 can be genetically separated from its inhibitory effect on cell cycle entry. EMBO J. 16: 4628–4638
Spaulding SW . 1993 The ways in which hormones change cyclic adenosine 3′,5′-monophosphate-dependent protein kinase subunits, and how such changes affect cell behavior. Endocr. Rev. 14: 632–650
Gjertsen B and Doskeland SO . 1995 Protein phosphorylation in apoptosis. Biochim. Biophys. Acta 1269: 187–199
Dumont J, Jauniaux JC and Roger PP . 1989 The cyclic AMP-mediated stimulation of cell proliferation. Trends Biochem. Sci. 14: 67–71
Braumann T, Jastorff B and Richter-Landsberg C . 1986 Fate of cyclic nucleotides in PC12 cell cultures: uptake, metabolism, and effects of metabolites on nerve growth factor-induced neurite outgrowth. J. Neurochem. 47: 912–919
Nicolai S, Willems J, Zwijsen A, Van Mechelen E and Slegers H . 1996 Cyclic AMP-induced differentiation increases the synthesis of extracellular superoxide dismutase in rat C6 glioma. Free Radic. Biol. Med. 21: 481–486
Sharma SK and Raj AB . 1987 Transient increase in intracellular concentration of adenosine 3′ : 5′-cyclic monophosphate results in morphological and biochemical differentiation of C6 glioma cells in culture. J. Neurosci. Res. 17: 135–141
Ginty DD, Fanger GR, Wagner JA and Maue RA . 1992 The activity of cAMP-dependent protein kinase is required at a posttranslational level for induction of voltage-dependent sodium channels by peptide growth factors in PC12 cells. J. Cell Biol. 116: 1465–1473
Kvanta A and Fredholm BB . 1993 Synergistic effects between protein kinase C and cAMP on activator protein-1 activity and differentiation of PC-12 pheocyromocytoma cells. J. Mol. Neurosci. 4: 205–214
McConkey DJ, Orrenius S, Okret S and Jondal M . 1993 Cyclic AMP potentiates glucocorticoid-induced endogenous endonuclease activation in thymocytes. FASEB J. 7: 580–588
Brown D and Phipps RP . 1996 Bcl-2 expression inhibits prostaglandin E2-mediated apoptosis in B cell lymphomas. J. Immunol. 157: 1359–1370
Suzuki K, Kizaki H, Tadakuma T and Ishimura Y. . 1990 12-O-tetradecanoylphorbol 13-acetate potentiates the action of cAMP in inducing DNA cleavage in thymocytes. Biochem. Biophys. Res. Commun. 171: 827–831
Gruol D and Altschmied J . 1993 Synergistic induction of apoptosis with glucocorticoids and 3′, 5′-cyclic adenosine monophosphate reveals agonist activity RU 486. Mol. Endocrinol. 7: 104–113
Dowd D and Miesfeld RL . 1992 Evidence that glucocorticoid- and cyclic AMP-induced apoptotic pathways in lymphocytes share distal events. Mol. Cell Biol. 12: 3600–3608
Parvathenami LK, Buescher ES, Chacon-Cruz E and Beebe SJ . 1998 Type 1 cAMP-dependent protein kinase delays apoptosis in Human Neutrophils at a site upstream of caspase-3. J. Biol. Chem. 273: 6736–6743
Ottonello L, Gonella R, Dapino P, Sacchetti C and Dallegri F . 1998 Prostaglandin E2 inhibits apoptosis in human neutrophilic polymorphonuclear leukocytes: Role of intracellular cyclic AMP levels. Exp. Hematol. 26: 895–902
Rossi AG, Cousin JM, Dransfield I, Lawson MF, Chilvers ER and Haslett C . 1995 Agents that elevate cAMP inhibit human neutrophil apoptosis. Biochem. Biophys. Res. Commun. 217: 892–899
Lomo J, Blomhoff HK, Beiske K, Stokke T and Smeland EB . 1995 TGF-beta 1 and cyclic AMP promote apoptosis in resting human B lymphocytes. J. Immunol. 154: 1634–1643
Goldstein B, Rogelj S, Siegel S, Farmer SR and Niles RM . 1990 Cyclic adenosine monophosphate-mediated induction of F9 teratocarcinoma differentiation in the absence of retinoic acid. J. Cell Physiol. 143: 205–212
Martin CA, Ziegler LM and Napoli JL . 1990 Retinoic acid, dibutyryl-cAMP, and differentiation affect the expression of retinoic acid receptors in F9 cells. Proc. Natl. Acad. Sci. U.S.A. 87: 4804–4808
Damjanov I, Zhu ZM, Andrews PW and Fenderson BA . 1994 Embryonal carcinoma cells differentiate into parietal endoderm via an intermediate stage corresponding to primitive endoderm. In Vivo 8: 967–973
Taneja R, Rochette-Egly C, Plassat JL, Penna L, Gaub MP and Chambon P . 1997 Phosphorylation of activation functions AF-1 and AF-2 of RAR alpha and RAR gamma is indispensable for differentiation of F9 cells upon retinoic acid and cAMP treatment. EMBO J. 16: 6452–6465
Olsson IL, Breitman TR and Gallo RC . 1982 Priming of human myeloid leukemic cell lines HL-60 and U-937 with retinoic acid for differentiation effects of cyclic adenosine 3′ : 5′-monophosphate-inducing agents and a T-lymphocyte-derived differentiation factor. Cancer Res. 42: 3928–3933
Ruchaud S, Duprez E, Gendron MC, Houge G, Genieser HG, Jastorff B, Doskeland SO and Lanotte M . 1994 Two distinctly regulated events, priming and triggering, during retinoid-induced maturation and resistance of NB4 promyleocytic leukemia cell line. Proc. Natl. acad. Sci. U.S.A. 91: 8428–8432
Gianni M, Terao M, Norio P, Barbui T, Rambaldi A and Garattini E . 1995 All-trans retinoic acid and cyclic adenosine monophosphate cooperate in the expression of leukocyte alkaline phosphatase in acute promyleocytic leukemia cells. Blood 85: 3619–3635
Chen A, Licht JD, Wu Y, Hellinger N, Scher W and Waxman S . 1994 Retinoic acid is required for and potentiates differentiation of acute promyelocytic leukemia cells by nonretinoid agents. Blood 84: 2122–2129
Quenech'Du N, Ruchaud S, Khelef N, Guiso N and Lanotte M . 1998 A sustained increase in the endogenous level of cAMP reduces the retinoid concentration required for APL cell maturation to near physiological levels. Leukemia 12: 1829–1833
Benoit G, Altucci L, Flexor M, Ruchaud S, Lillehaug J, Raffelsberger W, Gronemeyer H and Lanotte M . 1999 RAR-independent RXR signaling induces t(15;17) leukemia cell maturation. EMBO J. 18: 7011–7018
Lanotte M, Hermouet S, Gombaud-Saintonge G and Dobo I . 1986 On growth regulation of the rat promyelocytic leukemia (BNML): growth inhibition and eradication of clonogenic cells by cholera toxin. Leuk. Res. 10: 1319–1326
Gjertsen BT, Cressey LI, Ruchaud S, Houge G, Lanotte M and Doskeland SO . 1994 Multiple apoptotic death types triggered through activation of separate pathways by cAMP and inhibitors of protein phosphatases in one (IPC leukemia) cell line. J. Cell Sci. 107: 3363–3377
Lanotte M, Riviere JB, Hermouet S, Houge G, Vintermyr OK, Gjertsen BT and Doskeland SO . 1991 Programmed cell death (apoptosis) is induced rapidly and with positive cooperativity by activation of cyclic adenosine monophosphate-kinase I in a myeloid leukemia cell line. J. Cell Physiol. 146: 73–80
Vintermyr O, Gjertsen BT, Lanotte M and Doskeland SO . 1993 Microinjected catalytic subunit of cAMP-dependent protein kinase induces apoptosis in myeloid leukemia (IPC-81) cells. Exp. Cell. Res. 206: 157–161
Ruchaud S, Zorn M, Winkler E, Genieser HG, Doskeland SO, Jastorff B and Lanotte M . 1995 Evidence for several pathways of biological responses to hydrolysable cAMP-analogs using a model system of apoptosis. Cell. Pharmacol. 2: 127–140
Ruchaud S, Seite P, Foulkes NS, Sassone-Corsi P and Lanotte M . 1997 The transcriptional repressor ICER and cAMP-induced programmed cell death. Oncogene 15: 827–836
Martens A, Van Bekkum DW and Hagenbeek A . 1990 The BN acute myelocytic leukemia (BNML) (a rat model for studying human acute myelocytic leukemia (AML)). Leukemia 4: 241–257
Hawley RG, Lieu FH, Fong AZ and Hawley TS . 1994 Versatile retroviral vectors for potential use in gene therapy. Gene Ther. 1: 136–138
Pear WS, Nolan GP, Scott ML and Baltimore D . 1993 Production of high-titer helper-free retroviruses by transient transfection. Proc. Natl. Acad. Sci. U.S.A. 90: 8392–8396
Duprez E, Gjertsen BT, Bernard O, Lanotte M and Doskeland SO . 1993 Antiapoptotic effect of heterozygously expressed mutant RI (Ala336→Asp) subunit of cAMP kinase I in a rat leukemia cell line. J. Biol. Chem. 268: 8332–8340
Darzynkiewicz Z, Bruno S, Del Bino G, Gorczyca W, Hotz MA, Lassota P and Traganos F . 1992 Features of apoptotic cells measured by flow cytometry. Cytometry 13: 795–808
Jagels MA and Hugli TE . 1994 Mechanisms and mediators of neutrophilic leucocytosis. Immunopharmacology 28: 1–18
Savill J and Haslett C . 1995 Granulocyte clearance by apoptosis in the resolution of inflammation. Semin. Cell Biol. 6: 385–393
Coffey RG . 1992 Effects of cyclic nucleotides on granulocytes. Immunol. Ser. 57: 301–338
Ongini E and Fredholm BB . 1996 Pharmacology of adenosine A2A receptors. Trends Pharmacol. Sci. 17: 364–372
Elferink J and VanUffelen BE . 1996 The role of cyclic nucleotides in neutrophil migration. Gen. Pharmacol. 27: 387–393
Sigurdsson F, Khanna-Gupta A, Lawson N and Berliner N . 1997 Control of late neutrophil-specific gene expression: insights into regulation of myeloid differentiation. Semin. Hematol. 34: 303–310
Borner C . 1996 Diminished cell proliferation associated with the death-protective activity of Bcl-2. J. Biol. Chem. 271: 12695–12698
Vairo G, Innes KM and Adams JM . 1996 Bcl-2 has a cell cycle inhibitory function separable from its enhancement of cell survival. Oncogene 13: 1511–1519
Park J, Robertson K, Hickstein DD, Tsai S, Hockenbery DM and Collins SJ . 1994 Dysregulated bcl-2 expression inhibits apoptosis but not differentiation of retinoic acid-induced HL-60 granulocytes. Blood 84: 440–445
Lagasse E and Weissman IL . 1994 Bcl-2 inhibits Apoptosis of neutrophils but not their engulfment by macrophages. J. Exp. Med. 179: 1047–1052
Fairbairn LJ, Cowling GJ, Reipert BM and Dexter TM . 1993 Suppression of apoptosis allows differentiation and development of a multipotent hemopoietic cell line in the absence of added growth factors. Cell 74: 823–832
Maniatis T, Fritsch EF and Sambrook J . 1989 Molecular Cloning, A Laboratory Manual. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, NY.
Hillion J, Le Coniat M, Jonveaux P, Berger R and Bernard OA . 1997 AF6q21, a novel partner of the MLL gene in t(6;11)(q21;q23), defines a forkhead transcriptional factor subfamily. Blood 90: 3714–3719
Chomczynski P and Sacchi N . 1987 Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162: 156–159
Williams G . 1987 High-efficiency preparation of DNA from limiting quantities of eukaryotic cells for hybridization analysis. Gene 53: 121–126
Wright S, Schellenberger U, Wang H, Wang Y and Kinder D . 1998 Chemotherapeutic drug activation of the AP24 protease in apoptosis: requirement for caspase-3-like-proteases. Biochem. Biophys. Res. Commun. 245: 797–803
Acknowledgements
P Séité and S Ruchaud have contributed equally to the achievement of this work. Authors wish to thank Dr. T McDonnell (MD Anderson Cancer Institute, Houston, TX) for providing us with human Bcl-2 cDNA. S Ruchaud was supported by a grant from the Ligue Nationale contre le Cancer. This work has benefited from EU grants (TMR program), Ligue Nationale contre le Cancer, Ligue contre le Cancer des Yvelines, Fondation de France, Association pour la Recherche contre le Cancer and the Norwegian Research Council.
Author information
Authors and Affiliations
Corresponding author
Additional information
Edited by J Savill
Rights and permissions
About this article
Cite this article
Séité, P., Ruchaud, S., Hillion, J. et al. Ectopic expression of Bcl-2 switches over nuclear signalling for cAMP-induced apoptosis to granulocytic differentiation. Cell Death Differ 7, 1081–1089 (2000). https://doi.org/10.1038/sj.cdd.4400730
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/sj.cdd.4400730
Keywords
This article is cited by
-
Anti-microbial and cytotoxic 1,6-dihydroxyphenazine-5,10-dioxide (iodinin) produced by Streptosporangium sp. DSM 45942 isolated from the fjord sediment
Applied Microbiology and Biotechnology (2014)
-
Cyclic AMP induces IPC leukemia cell apoptosis via CRE-and CDK-dependent Bim transcription
Cell Death & Disease (2011)
-
Camptothecin and khat (Catha edulis Forsk.) induced distinct cell death phenotypes involving modulation of c-FLIPL, Mcl-1, procaspase-8 and mitochondrial function in acute myeloid leukemia cell lines
Molecular Cancer (2009)
-
Gap junctions and the propagation of cell survival and cell death signals
Apoptosis (2005)
-
Differentiation, Apoptosis, and Function of Human Immature and Mature Myeloid Cells: Intracellular Signaling Mechanism
International Journal of Hematology (2001)
