International Journal of Hematology

, Volume 92, Issue 1, pp 129–135 | Cite as

Targeting Smad4 links microRNA-146a to the TGF-β pathway during retinoid acid induction in acute promyelocytic leukemia cell line

  • Hua Zhong
  • Hai-rong Wang
  • Shuang Yang
  • Ji-hua Zhong
  • Ting Wang
  • Chun Wang
  • Fang-yuan Chen
Original Article


The expression pattern of microRNAs (miRNAs) and their potential target genes were investigated in acute promyelocytic leukemia (APL) cell line NB4 cells during all-trans-retinoid acid (ATRA) treatment by using a miRNA microarrays platform and real-time quantitative PCR (RTQ-PCR). MiR-146a as one of the miRNAs down-regulated by ATRA during APL differentiation was identified. Direct interaction between miR146a and its predictive target gene Smad4 were confirmed by Luciferase assay. Down-regulation of miR-146a and upregulation of Smad4 at protein levels were demonstrated. These data suggested that miR-146a might influence proliferation of APL cells through TGF-β1/Smad signal transduction pathway during ATRA induction.


miRNA Acute promyelocytic leukemia TGFβ/Smad signal transduction pathway Smad4 


  1. 1.
    Massagué J, Blain SW, Lo RS. TGFbeta signaling in growth control, cancer, and heritable disorders. Cell. 2000;103(2):295–309.CrossRefPubMedGoogle Scholar
  2. 2.
    Isufi I, Seetharam M, Zhou L, et al. Transforming growth factor-beta signaling in normal and malignant hematopoiesis. J Interferon Cytokine Res. 2007;27(7):543–52.CrossRefPubMedGoogle Scholar
  3. 3.
    Shi Y, Massague J. Mechanisms of TGF-beta signaling from cell membrane to the nucleus. Cell. 2003;113(6):685–700.CrossRefPubMedGoogle Scholar
  4. 4.
    Imai Y, Kurokawa M, Izutsu K, et al. Mutation of the Smad4 gene in acute myelogeneous leukemia and their functional implications in leukemogenesis. Oncogene. 2001;20:88–96.CrossRefPubMedGoogle Scholar
  5. 5.
    Zhang Y, Derynck R. Regulation of Smad signalling by protein associations and signalling crosstalk. Trends Cell Biol. 1999;9(7):274–9.CrossRefPubMedGoogle Scholar
  6. 6.
    Shih HM, Chang CC, Kuo HY, Lin DY. Daxx mediates SUMO-dependent transcriptional control and subnuclear compartmentalization. Biochem Soc Trans. 2007;35(Pt 6):1397–400.CrossRefPubMedGoogle Scholar
  7. 7.
    Hackanson B, Bennett KL, Brena RM, et al. Epigenetic modification of CCAAT/enhancer binding protein alpha expression in acute myeloid leukemia. Cancer Res. 2008;68(9):3142–51.CrossRefPubMedGoogle Scholar
  8. 8.
    Croce CM, Calin GA. miRNAs, cancer, and stem cell division. Cell. 2005;122(1):6–7.CrossRefPubMedGoogle Scholar
  9. 9.
    Kluiver J, Kroesen B-J, Poppema S, et al. The role of microRNAs in normal hematopoiesis and hematopoietic malignancies. Leukemia. 2006;20:1931–6.CrossRefPubMedGoogle Scholar
  10. 10.
    Garzon R, Pichiorri F, Palumbo T, et al. MicroRNA gene expression during retinoic acid-induced differentiation of human acute promyelocytic leukemia. Oncogene. 2007;26:4148–57.CrossRefPubMedGoogle Scholar
  11. 11.
    Da MW. A preliminary report of a quick method for determining drug sensitivity of leukemic cells in vitro. Zhonghua Nei Ke Za Zhi. 1982;21(2):109–12.PubMedGoogle Scholar
  12. 12.
    Chen C, Ridzon DA, Broomer AJ, et al. Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res. 2005;33:e179.CrossRefPubMedGoogle Scholar
  13. 13.
    Saumet A, Vetter G, Bouttier M, et al. Transcriptional repression of microRNA genes by PML-RARA increases expression of key cancer proteins in acute promyelocytic leukemia. Blood. 2009;113(2):412–21.CrossRefPubMedGoogle Scholar
  14. 14.
    Isken F, Steffen B, Merk S, et al. Identification of acute myeloid leukaemia associated microRNA expression patterns. Br J Haematol. 2008;140(2):153–61.CrossRefPubMedGoogle Scholar
  15. 15.
    Garzon R, Volinia S, Liu CG, et al. MicroRNA signatures associated with cytogenetics and prognosis in acute myeloid leukemia. Blood. 2008;111(6):3183–9.CrossRefPubMedGoogle Scholar
  16. 16.
    John B, Enright AJ, Aravin A, Tuschl T, Sander C, Marks DS. Human microRNA targets. PLoS Biol. 2004;2(11):1862–79.CrossRefGoogle Scholar
  17. 17.
    Lewis BP, Shih IH, Jones-Rhoades MW, Bartel DP, Burge CB. Prediction of mammalian microRNA targets. Cell. 2003;115(7):787–98.CrossRefPubMedGoogle Scholar
  18. 18.
    Fazi F, Rosa A, Fatica A, et al. A minicircuitry comprised of microRNA-223 and transcription factors NFI-A and C/EBPalpha regulates human granulopoiesis. Cell. 2005;123(5):819–31.CrossRefPubMedGoogle Scholar
  19. 19.
    Jongen-Lavrencic M, Sun SM, Dijkstra MK, Valk PJ, Löwenberg B. MicroRNA expression profiling in relation to the genetic heterogeneity of acute myeloid leukemia. Blood. 2008;111(10):5078–85.CrossRefPubMedGoogle Scholar
  20. 20.
    Debernardi S, Skoulakis S, Molloy G, Chaplin T, Dixon-McIver A, Young BD. MicroRNA miR-181a correlates with morphological sub-class of acute myeloid leukaemia and the expression of its target genes in global genome-wide analysis. Leukemia. 2007;21(5):912–6.PubMedGoogle Scholar
  21. 21.
    Zhong H, Chen FY, Wang HR, Lin JY, Xu R, Zhong JH, Huang HH. Modification of TGF-beta1 signaling pathway during NB4 cells differentiation by all-trans retinoid acid induction. Int J Hematol. 2009;89(4):438–44.CrossRefPubMedGoogle Scholar
  22. 22.
    Imai Y, Kurokawa M, Izutsu K, Hangaishi A, Maki K, Ogawa S, Chiba S, Mitani K, Hirai H. Mutations of the Smad4 gene in acute myelogeneous leukemia and their functional implications in leukemogenesis. Oncogene. 2001;20(1):88–96.CrossRefPubMedGoogle Scholar
  23. 23.
    Fatica A, Rosa A, Ballarino M, De Marchis ML, Rasmussen KD, Bozzoni I. Role of microRNAs in myeloid differentiation. Biochem Soc Trans. 2008;36(Pt 6):1201–5.CrossRefPubMedGoogle Scholar
  24. 24.
    De Marchis ML, Ballarino M, Salvatori B, Puzzolo MC, Bozzoni I, Fatica A. A new molecular network comprising PU.1, interferon regulatory factor proteins and miR-342 stimulates ATRA-mediated granulocytic differentiation of acute promyelocytic leukemia cells. Leukemia. 2009;23(5):856–62.CrossRefPubMedGoogle Scholar

Copyright information

© The Japanese Society of Hematology 2010

Authors and Affiliations

  • Hua Zhong
    • 1
  • Hai-rong Wang
    • 1
  • Shuang Yang
    • 1
  • Ji-hua Zhong
    • 1
  • Ting Wang
    • 1
  • Chun Wang
    • 2
  • Fang-yuan Chen
    • 1
  1. 1.Department of Hematology, Renji HospitalShanghai Jiaotong University, School of MedicineShanghaiPeople’s Republic of China
  2. 2.Department of Hematology, Shanghai First People’s HospitalShanghai Jiaotong UniversityShanghaiPeople’s Republic of China

Personalised recommendations