Skip to main content

Promoted megakaryocytic differentiation of K562 cells through oxidative stress caused by near ultraviolet irradiation

Abstract

Reactive oxygen species (ROS) have been proven to be important activators for various cellular activities, including cell differentiation. Several reports showed the necessity of ROS during cell differentiation of the megakaryocytic (MK) lineage. In this study, we employed near ultraviolet (near-UV) irradiation to generate endogenous oxidative stress in an MK differentiation process of K562 cells with phorbol 12-myristate 13-acetate (PMA) induction. A significant increase in the intracellular ROS level was detected on day 1 after near-UV irradiation. In the initial stage of differentiation, a shifted fraction of G1 and G2 phase cells was obtained using near-UV irradiation, giving an increased percentage of G2 phase cells (up from 31.1 to 68.7%). The near-UV irradiation-induced upregulation of the p21 gene, which is a cell cycle inhibitor, suggested that the G2 phase cells were prevented from undergoing cell division. It was found that the percentage of high ploidy (8N and 16N) cells was enhanced significantly at the later stage of the K562 cell culture with near-UV irradiation. Moreover, time-lapse analysis showed that near-UV irradiation encouraged the expression of CD41, a specific surface marker of megakaryocytes. This is the first report that the elevated oxidative stress through the near-UV irradiation promoted the MK differentiation of PMA-induced K562 cells.

Abbreviations

EDTA:

ethylene diamine tetra-acetic acid

MK:

megakaryocytic

PBS:

phosphate-buffered saline

PMA:

phorbol 12-myristate 13-acetate

ROS:

reactive oxygen species

References

  1. Halliwell, B. and Aruoma, O.I. DNA damage by oxygen-derived species. FEBS Lett. 28 (1991) 9–19.

    Article  Google Scholar 

  2. Gruijl, F.R., Kranen, H.J. and Mullenders, L.H.F. UV-induced DNA damage, repair, mutations and oncogenic pathways in skin cancer. J. Photochem. Photobiol. B: Biology 63 (2001) 19–27.

    Article  PubMed  Google Scholar 

  3. Halliwell, B. Free radicals, reactive oxygen species and human disease: a critical evaluation with special reference to atherosclerosis. Br. J. Exp. Path. 70 (1989) 737–757.

    CAS  Google Scholar 

  4. Sardina J.L., Lopez-Ruano, G., Sanchez-Abarca, L.I., Perez-Simon, J.A., Gaztelumendi, A., Trigueros, C., Llanillo, M., Sanchez-Yague, J. and Hernandez-Hernandez, A. p22phox-dependent NADPH oxidase activity is required for megakaryocytic differentiation. Cell Death Differ. 17 (2010) 1842–1854.

    Article  CAS  PubMed  Google Scholar 

  5. Mostafa, S.S., Miller, W.M. and Papoutsakis, E.T. Oxygen tension influences the differentiation, maturation and apoptosis of human megakaryocytes. Br. J. Haematol. 111 (2000) 879–889.

    CAS  PubMed  Google Scholar 

  6. Hirose, K., Monzen, S., Sato, H., Sato, M., Aoki, M., Hatayama, Y., Kawaguchi, H., Narita, Y., Takai, Y. and Kashiwakura, I. Megakaryocytic differentiation in human chronic myelogenous leukemia K562 cells induced by ionizing radiation in combination with phorbol 12-myristate 13-acetate. J. Radiat. Res. 54 (2013) 438–446.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  7. Ojima, Y., Duncan, M.T., Nurhayati, R.W., Taya, M. and Miller W.M. Synergistic effect of hydrogen peroxide on polyploidization during the megakaryocytic differentiation of K562 leukemia cells by PMA. Exp. Cell Res. 319 (2013) 2205–2215.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  8. Butler, T.M., Ziemiecki, A. and Friis, R.R. Megakaryocytic differentiation of K562 cells is associated with changes in the cytoskeletal organization and the pattern of chromatographically distinct forms of phosphotyrosyl-specific protein phosphatases. Cancer Res. 50 (1990) 6323–6329.

    CAS  PubMed  Google Scholar 

  9. Collamonici, O.R., Trepel, J.B., Vidal, C.A. and Neckers L.M. Phobol ester induces c-sis gene transcription in stem cell line K-562. Mol. Cell Biol. 6 (1986) 1847–1850.

    Google Scholar 

  10. Tetteroo, P.A., Massaro, F., Mulder, A., Schreuder van Gelder, R. and von dem Borne, A.E.G. Megakaryoblastic differentiation of proerythroblastic K562 cell-line cells. Leukem. Res. 8 (1984) 197–206.

    Article  CAS  Google Scholar 

  11. Fukuda, M. Tumor-promoting phorbol diester-induced specific changes in cell surface glycoprotein profile of K562 human leukemic cells. Cancer Res. 41 (1981) 4621–4628.

    CAS  PubMed  Google Scholar 

  12. Giammona, L.M., Fuhrken, P.G., Papoutsakis, E.T. and Miller, W.M. Nicotinamide (vitamin B3) increases the polyploidization and proplatelet formation of cultured primary human megakaryocytes. Br. J. Haematol. 135 (2006) 554–566.

    Article  CAS  PubMed  Google Scholar 

  13. Shelly, C., Petruzzelli, L. and Herrera, R. K562 cells resistant to phorbol 12-myristate 13-acetate-induced growth arrest: dissociation of mitogenactivated protein kinase activation and Egr-1 expression from megakaryocyte differentiation. Cell Death Differ. 11 (2000) 501–506.

    CAS  Google Scholar 

  14. Kulkarni, A.P., Mittal, S.P.K., Devasagayam, T.P.A. and Pal, J.K. Oxidative stress perturbs cell proliferation in human K562 cells by modulating protein synthesis and cell cycle. Free Rad. Res. 43 (2009) 1090–1100.

    Article  CAS  Google Scholar 

  15. Nagata, Y., Muro, Y. and Todokoro, K. Thrombopoietin-induced polyploidization of bone marrow megakaryocytes is due to a unique regulatory mechanism in late mitosis. J. Cell Biol. 139 (1997) 449–456.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  16. Baccini, V., Roy, L., Vitrat, N., Chagraoui, H., Sabri, S., Couedic, J.P., Debili, N., Wendling, F. and Vainchenker, W. Role of p21Cip1/Waf1 in cellcycle exit of endomitotic megakaryocytes. Blood 98 (2001) 3274–3282.

    Article  CAS  PubMed  Google Scholar 

  17. Wuerzerberger-Davis, S.M., Chang, P.Y., Berchtold, C. and Miyamoto, S. Enhanced G2-M arrest by nuclear factor-κ-B-dependent p21waf1/cip1 induction. Mol. Cancer Res. 3 (2005) 345–352.

    Article  Google Scholar 

  18. Kang, C.D., Han, C.S., Kim, K.W., Do, I.R., Kim, C.M., Kim, S.H., Lee, E.Y. and Chung, B.S. Activation of NF-κB mediates the PMA-induced differentiation of K562 cells. Cancer Lett. 132 (1998) 99–106.

    Article  CAS  PubMed  Google Scholar 

  19. Mitjavila-Garcia, M.T., Cailleret, M., Godin, I., Nogueira, M.M., Cohen-Solal, K., Schiavon, V., Lecluse, Y., Pesteur, F., Lagrue, A.H. and Vainchenker, W. Expression of CD41 on hematopoietic progenitors derived from embryonic hematopoietic cells. Development 129 (2002) 2003–2013.

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Masahito Taya.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Nurhayati, R.W., Ojima, Y., Nomura, N. et al. Promoted megakaryocytic differentiation of K562 cells through oxidative stress caused by near ultraviolet irradiation. Cell Mol Biol Lett 19, 590–600 (2014). https://doi.org/10.2478/s11658-014-0215-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2478/s11658-014-0215-3

Keywords

  • Near ultraviolet
  • Irradiation
  • K562 cells
  • Megakaryocytic differentiation
  • Polyploidization
  • Reactive oxygen species
  • Cell cycle
  • Phorbol 12-myristate 13-acetate