Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

C/EBPα and C/EBPɛ induce the monocytic differentiation of myelomonocytic cells with the MLL-chimeric fusion gene

Abstract

CCAAT/enhancer binding proteins (C/EBPs) have an important function in granulocytic differentiation, and are also involved in the leukemogenesis of acute myeloid leukemia (AML). Their involvement in myelomonocytic leukemia, however, is still unclear. Therefore, the expression and function of C/EBPs in myelomonocytic cells with MLL-fusion genes were investigated. Retinoic acid (RA) induced monocytic differentiation in the myelomonocytic cell lines with MLL-fusion genes, THP-1, MOLM-14 and HF-6 cells, accompanied by monocytic differentiation with the upregulation of C/EBPα and C/EBPɛ. Monocytic differentiation by RA treatment was confirmed in primary AML cells using a clonogenic assay. When the activity of C/EBPα or C/EBPɛ was introduced into HF-6 cells, their cellular growth was arrested through differentiation into monocytes with the concomitant marked downregulation of Myc. Cebpe mRNA was upregulated by the induction of C/EBPα-ER, but not vice versa, thus suggesting that C/EBPɛ may have an important function in the differentiation process. Introduction of Myc isoforms into HF-6 cells partially antagonized the C/EBPs effects. These findings suggest that the ectopic expression of C/EBPɛ, as well as C/EBPα, can induce the monocytic differentiation of myelomonocytic leukemic cells with MLL-fusion gene through the downregulation of Myc, thus providing insight into the development of novel therapeutic approaches.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  • Ayton PM, Cleary ML . (2003). Transformation of myeloid progenitors by MLL oncoproteins is dependent on Hoxa7 and Hoxa9. Genes Dev 17: 2298–2307.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cozzio A, Passegue E, Ayton PM, Karsunky H, Cleary ML, Weissman IL . (2003). Similar MLL-associated leukemias arising from self-renewing stem cells and short-lived myeloid progenitors. Genes Dev 17: 3029–3035.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dahl R, Walsh JC, Lancki D, Laslo P, Iyer SR, Singh H et al. (2003). Regulation of macrophage and neutrophil cell fates by the PU.1 C/EBPalpha ratio and granulocyte colony-stimulating factor. Nat Immunol 4: 1029–1036.

    Article  CAS  PubMed  Google Scholar 

  • De Braekeleer M, Morel F, Le Bris MJ, Herry A, Douet-Guilbert N . (2005). The MLL gene and translocations involving chromosomal band 11q23 in acute leukemia. Anticancer Res 25: 1931–1944.

    CAS  PubMed  Google Scholar 

  • Freytag SO, Geddes TJ . (1992). Reciprocal regulation of adipogenesis by Myc and C/EBP alpha. Science 256: 379–382.

    Article  CAS  PubMed  Google Scholar 

  • Fukuchi Y, Shibata F, Ito M, Goto-Koshino Y, Sotomaru Y, Ito M et al. (2006). Comprehensive analysis of myeloid lineage conversion using mice expressing an inducible form of C/EBP alpha. EMBO J 25: 3398–3410.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gery S, Gombart AF, Fung YK, Koeffler HP . (2004). C/EBPepsilon interacts with retinoblastoma and E2F1 during granulopoiesis. Blood 103: 828–835.

    Article  CAS  PubMed  Google Scholar 

  • Gombart AF, Hofmann WK, Kawano S, Takeuchi S, Krug U, Kwok SH et al. (2002). Mutations in the gene encoding the transcription factor CCAAT/enhancer binding protein alpha in myelodysplastic syndromes and acute myeloid leukemias. Blood 99: 1332–1340.

    Article  CAS  PubMed  Google Scholar 

  • Gombart AF, Kwok SH, Anderson KL, Yamaguchi Y, Torbett BE, Koeffler HP . (2003). Regulation of neutrophil and eosinophil secondary granule gene expression by transcription factors C/EBP epsilon and PU.1. Blood 101: 3265–3273.

    Article  CAS  PubMed  Google Scholar 

  • Hemmi H, Breitman TR . (1985). Induction of functional differentiation of a human monocytic leukemia cell line (THP-1) by retinoic acid and cholera toxin. Jpn J Cancer Res 76: 345–351.

    CAS  PubMed  Google Scholar 

  • Iijima K, Honma Y, Niitsu N . (2004). Granulocytic differentiation of leukemic cells with t(9;11)(p22;q23) induced by all-trans-retinoic acid. Leuk Lymphoma 45: 1017–1024.

    Article  CAS  PubMed  Google Scholar 

  • Johansen LM, Iwama A, Lodie TA, Sasaki K, Felsher DW, Golub TR et al. (2001). c-Myc is a critical target for c/EBPalpha in granulopoiesis. Mol Cell Biol 21: 3789–3806.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kawada H, Ando K, Tsuji T, Shimakura Y, Nakamura Y, Chargui J et al. (1999). Rapid ex vivo expansion of human umbilical cord hematopoietic progenitors using a novel culture system. Exp Hematol 27: 904–915.

    Article  CAS  PubMed  Google Scholar 

  • Kitamura T, Koshino Y, Shibata F, Oki T, Nakajima H, Nosaka T et al. (2003). Retrovirus-mediated gene transfer and expression cloning: powerful tools in functional genomics. Exp Hematol 31: 1007–1014.

    Article  CAS  PubMed  Google Scholar 

  • Kubota T, Kawano S, Chih DY, Hisatake Y, Chumakov AM, Taguchi H et al. (2000). Representational difference analysis using myeloid cells from C/EBP epsilon deletional mice. Blood 96: 3953–3957.

    CAS  PubMed  Google Scholar 

  • Kummalue T, Friedman AD . (2003). Cross-talk between regulators of myeloid development: C/EBPalpha binds and activates the promoter of the PU.1 gene. J Leukoc Biol 74: 464–470.

    Article  CAS  PubMed  Google Scholar 

  • Lee IH, Lee JH, Lee MJ, Lee SY, Kim IS . (2002). Involvement of CCAAT/enhancer-binding protein alpha in haptoglobin gene expression by all-trans-retinoic acid. Biochem Biophys Res Commun 294: 956–961.

    Article  CAS  PubMed  Google Scholar 

  • Lee YJ, Jones LC, Timchenko NA, Perrotti D, Tenen DG, Kogan SC . (2006). CCAAT/enhancer binding proteins alpha and epsilon cooperate with all-trans retinoic acid in therapy but differ in their antileukemic activities. Blood 108: 2416–2419.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li ZY, Liu DP, Liang CC . (2005). New insight into the molecular mechanisms of MLL-associated leukemia. Leukemia 19: 183–190.

    Article  PubMed  Google Scholar 

  • Mueller BU, Pabst T, Fos J, Petkovic V, Fey MF, Asou N et al. (2006). ATRA resolves the differentiation block in t(15;17) acute myeloid leukemia by restoring PU. 1 expression. Blood 107: 3330–3338.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nakajima H, Ihle JN . (2001). Granulocyte colony-stimulating factor regulates myeloid differentiation through CCAAT/enhancer-binding protein epsilon. Blood 98: 897–905.

    Article  CAS  PubMed  Google Scholar 

  • Nakajima H, Watanabe N, Shibata F, Kitamura T, Ikeda Y, Handa M . (2006). N-terminal region of CCAAT/enhancer-binding protein epsilon is critical for cell cycle arrest, apoptosis, and functional maturation during myeloid differentiation. J Biol Chem 281: 14494–14502.

    Article  CAS  PubMed  Google Scholar 

  • Oelgeschlager M, Nuchprayoon I, Luscher B, Friedman AD . (1996). C/EBP, c-Myb, and PU.1 cooperate to regulate the neutrophil elastase promoter. Mol Cell Biol 16: 4717–4725.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ono R, Nakajima H, Ozaki K, Kumagai H, Kawashima T, Taki T et al. (2005). Dimerization of MLL fusion proteins and FLT3 activation synergize to induce multiple-lineage leukemogenesis. J Clin Invest 115: 919–929.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pabst T, Mueller BU, Harakawa N, Schoch C, Haferlach T, Behre G et al. (2001b). AML1-ETO downregulates the granulocytic differentiation factor C/EBPalpha in t(8;21) myeloid leukemia. Nat Med 7: 444–451.

    Article  CAS  PubMed  Google Scholar 

  • Pabst T, Mueller BU, Zhang P, Radomska HS, Narravula S, Schnittger S et al. (2001a). Dominant-negative mutations of CEBPA, encoding CCAAT/enhancer binding protein-alpha (C/EBPalpha), in acute myeloid leukemia. Nat Genet 27: 263–270.

    Article  CAS  PubMed  Google Scholar 

  • Pan Z, Hetherington CJ, Zhang DE . (1999). CCAAT/enhancer-binding protein activates the CD14 promoter and mediates transforming growth factor beta signaling in monocyte development. J Biol Chem 274: 23242–23248.

    Article  CAS  PubMed  Google Scholar 

  • Radomska HS, Basseres DS, Zheng R, Zhang P, Dayaram T, Yamamoto Y et al. (2006). Block of C/EBP alpha function by phosphorylation in acute myeloid leukemia with FLT3 activating mutations. J Exp Med 203: 371–381.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Schoch C, Schnittger S, Klaus M, Kern W, Hiddemann W, Haferlach T . (2003). AML with 11q23/MLL abnormalities as defined by the WHO classification: incidence, partner chromosomes, FAB subtype, age distribution, and prognostic impact in an unselected series of 1897 cytogenetically analyzed AML cases. Blood 102: 2395–2402.

    Article  CAS  PubMed  Google Scholar 

  • Schreiner S, Birke M, Garcia-Cuellar MP, Zilles O, Greil J, Slany RK . (2001). MLL-ENL causes a reversible and myc-dependent block of myelomonocytic cell differentiation. Cancer Res 61: 6480–6486.

    CAS  PubMed  Google Scholar 

  • Schuster MB, Porse BT . (2006). C/EBPalpha: a tumour suppressor in multiple tissues? Biochim Biophys Acta 1766: 88–103.

    CAS  PubMed  Google Scholar 

  • Tavor S, Vuong PT, Park DJ, Gombart AF, Cohen AH, Koeffler HP . (2002). Macrophage functional maturation and cytokine production are impaired in C/EBP epsilon-deficient mice. Blood 99: 1794–1801.

    Article  CAS  PubMed  Google Scholar 

  • Truong BT, Lee YJ, Lodie TA, Park DJ, Perrotti D, Watanabe N et al. (2003). CCAAT/enhancer binding proteins repress the leukemic phenotype of acute myeloid leukemia. Blood 101: 1141–1148.

    Article  CAS  PubMed  Google Scholar 

  • Walkley CR, Purton LE, Snelling HJ, Yuan YD, Nakajima H, Chambon P et al. (2004). Identification of the molecular requirements for an RAR alpha-mediated cell cycle arrest during granulocytic differentiation. Blood 103: 1286–1295.

    Article  CAS  PubMed  Google Scholar 

  • Wang D, D’Costa J, Civin CI, Friedman AD . (2006). C/EBPalpha directs monocytic commitment of primary myeloid progenitors. Blood 108: 1223–1229.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Wang X, Scott E, Sawyers CL, Friedman AD . (1999). C/EBPalpha bypasses granulocyte colony-stimulating factor signals to rapidly induce PU.1 gene expression, stimulate granulocytic differentiation, and limit proliferation in 32D cl3 myeloblasts. Blood 94: 560–571.

    CAS  PubMed  Google Scholar 

  • Williams SC, Du Y, Schwartz RC, Weiler SR, Ortiz M, Keller JR et al. (1998). C/EBPepsilon is a myeloid-specific activator of cytokine, chemokine, and macrophage-colony-stimulating factor receptor genes. J Biol Chem 273: 13493–13501.

    Article  CAS  PubMed  Google Scholar 

  • Yamanaka R, Barlow C, Lekstrom-Himes J, Castilla LH, Liu PP, Eckhaus M et al. (1997). Impaired granulopoiesis, myelodysplasia, and early lethality in CCAAT/enhancer binding protein epsilon-deficient mice. Proc Natl Acad Sci USA 94: 13187–13192.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang DE, Hetherington CJ, Meyers S, Rhoades KL, Larson CJ, Chen HM et al. (1996). CCAAT enhancer-binding protein (C/EBP) and AML1 (CBF alpha2) synergistically activate the macrophage colony-stimulating factor receptor promoter. Mol Cell Biol 16: 1231–1240.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang P, Iwasaki-Arai J, Iwasaki H, Fenyus ML, Dayaram T, Owens BM et al. (2004). Enhancement of hematopoietic stem cell repopulating capacity and self-renewal in the absence of the transcription factor C/EBP alpha. Immunity 21: 853–863.

    Article  CAS  PubMed  Google Scholar 

  • Zheng R, Friedman AD, Levis M, Li L, Weir EG, Small D . (2004). Internal tandem duplication mutation of FLT3 blocks myeloid differentiation through suppression of C/EBPalpha expression. Blood 103: 1883–1890.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Dr Toshio Kitamura (Institute of Medical Science, University of Tokyo, Tokyo, Japan) for providing PLAT-E cells. This study was supported in part by Research and Study Program of Tokai University Educational System General Research Organization and a Grant-in Aid for Scientific Research (C) No. 19591139 from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to H Matsushita.

Additional information

Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Matsushita, H., Nakajima, H., Nakamura, Y. et al. C/EBPα and C/EBPɛ induce the monocytic differentiation of myelomonocytic cells with the MLL-chimeric fusion gene. Oncogene 27, 6749–6760 (2008). https://doi.org/10.1038/onc.2008.285

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2008.285

Keywords

This article is cited by

Search

Quick links