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Human Cell

, Volume 28, Issue 2, pp 65–72 | Cite as

BMS-777607 promotes megakaryocytic differentiation and induces polyploidization in the CHRF-288-11 cells

  • Retno Wahyu Nurhayati
  • Yoshihiro Ojima
  • Masahito TayaEmail author
Research Article

Abstract

Introduction of a polyploidy inducer is a promising strategy to achieve a high level of polyploidization during megakaryocytic (MK) differentiation. Here, we report that a multi-kinase inhibitor, BMS-777607, is a potent polyploidy inducer for elevating high ploidy cell formation in the MK-differentiated CHRF-288-11 (CHRF) cells. Our result showed that BMS-777607 strongly inhibited cell division without affecting cell viability when detected at day 1 after treatment. As a consequence, the high ploidy (≥8N) cells were accumulated in culture for 8 days, with an increase from 16.2 to 75.2 % of the total cell population. The elevated polyploidization was accompanied by the increased expression level of MK marker, CD41 (platelet glycoprotein IIb/IIIa, GPIIb/IIIa), suggesting that BMS-777607 promoted both polyploidization and commitment of MK-differentiated CHRF cells. Platelet-like fragments (PFs) were released by mature CHRF cells. Based on a flow cytometry assay, it was found that the PFs produced from BMS-777607-treated cells tended to have larger size and higher expression of GPIIb/IIIa, a receptor for platelet adhesion. Taken together, these results suggested that BMS-777607 promoted MK differentiation of CHRF cells and increased the functional property of platelet-like fragments.

Keywords

Polyploidy Megakaryocytic differentiation CHRF-288-11 cells BMS-777607 Platelet-like fragment 

Notes

Acknowledgments

This research was in part supported by Grant-in-Aids for Scientific Researches, No. 25289295, from the Ministry of Education, Culture, Sports, Science and Technology of Japan. We thank Prof. William M. Miller of Northwestern University for kindly providing CHRF-288-11 and K562 cells.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

13577_2014_102_MOESM1_ESM.docx (115 kb)
Supplementary material 1 (DOCX 115 kb)

References

  1. 1.
    Tavassoli M. Megakaryocyte-platelet axis and the process of platelet formation and release. Blood. 1980;55(4):537–45.PubMedGoogle Scholar
  2. 2.
    Nagata Y, Muro Y, Todokoro K. Thrombopoietin-induced polyploidization of bone marrow megakaryocytes is due to a unique regulatory mechanism in late mitosis. J Cell Biol. 1997;139:449–57.CrossRefPubMedCentralPubMedGoogle Scholar
  3. 3.
    Lordier L, Jalil A, Aurade F, Larbret F, Larghero J, Debili N, Vainchenker W, Chang Y. Megakaryocyte endomitosis is a failure of late cytokinesis related to defects in the contractile ring and RHO/Rock signaling. Blood. 2008;112(8):3164–74.CrossRefPubMedGoogle Scholar
  4. 4.
    Avanzi MP, Chen A, He W, Mitchell WB. Optimizing megakaryocyte polyploidization by targeting multiple pathways of cytokinesis. Transfusion. 2012;52:2406–13.CrossRefPubMedGoogle Scholar
  5. 5.
    Giammona LM, Fuhrken PG, Papoutsakis ET, Miller WM. Nicotinamide (vitamin B3) increases the polyploidization and proplatelet formation of cultured primary human megakaryocytes. Br J Haematol. 2006;135:554–66.CrossRefPubMedGoogle Scholar
  6. 6.
    Lannuti BJ, Blake N, Gandhi MJ, Reems JA, Drachman JG. Induction of polyploidization in leukemic cell lines and primary bone marrow by Src kinase inhibitor SU6656. Blood. 2005;105(10):3875–8.CrossRefGoogle Scholar
  7. 7.
    Krause DS, Crispino JD. Molecular pathways: induction of polyploidy as a novel differentiation therapy for leukemia. Clin Cancer Res. 2013;19(22):6084–8.CrossRefPubMedGoogle Scholar
  8. 8.
    Schroeder GM, An Y, Cai ZW, Chen XT, Clark C, Cornelius LA, Dai J, Gullo-Brown J, Gupta A, Henley B, Hunt JT, Jeyaseelan R, Kamath A, Kim K, Lippy J, Lombardo LJ, Manne V, Oppenheimer S, Sack JS, Schmidt RJ, Shen G, Stefanski K, Tokarski JS, Trainor GL, Wautlet BS, Wei D, Williams DK, Zhang Y, Zhang Y, Fargnoli J, Borzilleri RM. Discovery of N-(4-(2-amino-3-chloropyridin-4-yloxy)-3-fluorophenyl)-4-ethoxy-1-(4-fluorophenyl)-2-oxo-1,2-dihydropyridine-3-carboxamide, BMS-777607, a selective and orally efficacious inhibitor of the Met kinase superfamily. J Med Chem. 2009;52(5):1251–4.CrossRefPubMedGoogle Scholar
  9. 9.
    Nigg EA. Mitotic kinases as regulators of cell division and its checkpoints. Nat Rev Mol Cell Biol. 2001;2(1):21–32.CrossRefPubMedGoogle Scholar
  10. 10.
    Sharma S, Zeng JY, Zhuang CM, Zhou YQ, Yao HP, Hu X, Zhang R, Wang MH. Small-molecule inhibitor BMS-777607 induces breast cancer cell polyploidy with increased resistance to cytotoxic chemotherapy agents. Mol Cancer Ther. 2013;12(5):725–36.CrossRefPubMedGoogle Scholar
  11. 11.
    Saito H. Megakaryocytic cell lines. Baillieres Clin Haemato. 1997;10(1):47–63.CrossRefGoogle Scholar
  12. 12.
    Fugman DA, Witte DP, Jones CL, Aronow BJ, Lieberman MA. In vitro establishment and characterization of a human megakaryoblastic cell line. Blood. 1990;75(6):1252–61.PubMedGoogle Scholar
  13. 13.
    Jiang F, Jia Y, Cohen I. Fibronectin- and protein kinase C-mediated activation of ERK/MAPK are essential for proplateletlike formation. Blood. 2002;99(10):3579–84.CrossRefPubMedGoogle Scholar
  14. 14.
    De Cuyper IM, Meinders M, van de Vijver E, de Korte D, Porcelijn L, de Haas M, Eble JA, Seeger K, Rutella S, Pagliara D, Kuijpers TW, Verhoeven AJ, van den Berg TK, Gutierrez L. A novel flow cytometry-based platelet aggregation assay. Blood. 2013;121(10):e70–80.CrossRefPubMedGoogle Scholar
  15. 15.
    Choi ES, Nichol JL, Hokom MM, Hornkohl AC, Hunt P. Platelets generated in vitro from proplatelet-displaying human megakaryocytes are functional. Blood. 1995;85(2):402–13.PubMedGoogle Scholar
  16. 16.
    Javela K, Kekomäki R. Mean platelet size related to glycoprotein-specific autoantibodies and platelet-associated IgG. Int J Lab Hematol. 2007;29(6):433–41.CrossRefPubMedCentralPubMedGoogle Scholar
  17. 17.
    Tovar C, Higgins B, Deo D, Kolinsky K, Liu J, Heimbrook DC, Vassilev LT. Small-molecule inducer of cancer cell polyploidy promotes apoptosis or senescence. Cell Cycle. 2010;9(16):3364–75.CrossRefPubMedGoogle Scholar
  18. 18.
    Ganem NJ, Pellman D. Limiting the proliferation of polyploid cells. Cell. 2007;131(3):437–40.CrossRefPubMedGoogle Scholar
  19. 19.
    Nakagawa M, Oliva JL, Kothapalli D, Fournier A, Assoian RK, Kazanietz MG. Phorbolester-induced G1 phase arrest selectively mediated by protein kinase Cδ-dependent induction of p21. J Biol Chem. 2005;280(40):33926–34.CrossRefPubMedGoogle Scholar
  20. 20.
    Yoshino T, Sakaguchi M, Masuda T, Kawakita M, Takatsuki K. Two regulation points for differentiation in the cell cycle of human megakaryocytes. Br J Haematol. 1996;92(4):780–7.CrossRefPubMedGoogle Scholar
  21. 21.
    Kosaka C, Sasaguri T, Ishida A, Ogata J. Cell cycle arrest in the G2 phase induced by phorbol ester and diacylglycerol in vascular endothelial cells. Am J Physiol. 1996;270:C170–8.PubMedGoogle Scholar
  22. 22.
    Apostolidis PA, Lindsey S, Miller WM, Papoutsakis ET. Proposed megakaryocytic regulon of p53: the genes engaged to control cell cycle and apoptosis during megakaryocytic differentiation. Physiol Genomics. 2012;44(12):638–50.CrossRefPubMedCentralPubMedGoogle Scholar
  23. 23.
    Limb JK, Song D, Jeon M, Han SY, Han G, Jhon GJ, Bae YS, Kim J. 2-(Trimethyl ammonium)ethyl (R)-3-methoxy-3-oxo-2-stearamidopropyl phosphate promotes megakaryocytic differentiation of myeloid leukaemia cells and primary human CD34+ haematopoietic stem cells. J Tissue Eng Regen Med. 2012;. doi: 10.1002/term.1628.PubMedGoogle Scholar
  24. 24.
    Mattia G, Vulcano F, Milazzo L, Barca A, Macioce G, Giampaolo A, Hassan HJ. Different ploidy levels of megakaryocytes generated from peripheral or cord blood CD34+ cells are correlated with different levels of platelet release. Blood. 2002;99(3):888–97.CrossRefPubMedGoogle Scholar
  25. 25.
    Zimmet J, Ravid K. Polyploidy: occurrence in nature, mechanisms, and significance for the megakaryocyte-platelet system. Exp Hematol. 2000;28(1):3–16.CrossRefPubMedGoogle Scholar
  26. 26.
    Bessman JD. The relation of megakaryocyte ploidy to platelet volume. Am J Hematol. 1984;16(2):161–70.CrossRefPubMedGoogle Scholar
  27. 27.
    Tomer A. Human marrow megakaryocyte differentiation: multiparameter correlative analysis identifies von Willebrand factor as a sensitive and distinctive marker for early (2N and 4N) megakaryocytes. Blood. 2004;104(9):2722–7.CrossRefPubMedGoogle Scholar
  28. 28.
    Lozzio CB, Lozzio BB. Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. Blood. 1975;45(3):321–34.PubMedGoogle Scholar
  29. 29.
    Tetteroo PA, Massaro F, Mulder A, Schreuder van Gelder R, von dem Borne AEG. Megakaryoblastic differentiation of proerythroblastic K562 cell-line cells. Leukem Res. 1984;8(2):197–206.CrossRefGoogle Scholar
  30. 30.
    Baliga BS, Mankad M, Shah AK, Mankad VN. Mechanism of differentiation of human erythroleukaemic cell line K562 by hemin. Cell Prolif. 1993;26(6):519–29.CrossRefPubMedGoogle Scholar
  31. 31.
    Kunicki TJ. Platelet membrane glycoproteins and their function: an overview. Blut. 1989;59(1):30–4.CrossRefPubMedGoogle Scholar
  32. 32.
    French DL, Seligsohn U. Platelet glycoprotein IIb/IIIa receptors and Glanzmann’s thrombasthenia. Arterioscler Thromb Vasc Biol. 2000;20(3):607–10.CrossRefPubMedGoogle Scholar
  33. 33.
    Eldor A, Avitzour M, Or R, Hanna R, Penchas S. Prediction of haemorrhagic diathesis in thrombocytopenia by mean platelet volume. Br Med J. 1982;285(6339):397–400.CrossRefGoogle Scholar
  34. 34.
    Martin JF, Trowbridge EA, Salmon G, Plumb J. The biological significance of platelet volume: its relationship to bleeding time, platelet thromboxane B2 production and megakaryocyte nuclear DNA concentration. Thromb Res. 1983;32(5):443–60.CrossRefPubMedGoogle Scholar

Copyright information

© Japan Human Cell Society and Springer Japan 2014

Authors and Affiliations

  • Retno Wahyu Nurhayati
    • 1
  • Yoshihiro Ojima
    • 1
  • Masahito Taya
    • 1
    Email author
  1. 1.Division of Chemical Engineering, Department of Materials Engineering Science, Graduate School of Engineering ScienceOsaka UniversityToyonakaJapan

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