Abstract
Exponentially growing human erythroleukemia K562 cells were permeabilized and the dose dependent decrease of DNA synthesis rate was measured after ultraviolet (UV B, 290 nm) irradiation. Cells were able to overcome 2 and 5 J/m2 UV doses, partial recovery was observed at 15 J/m2, while at high (25 J/m2) UV dose replicative DNA synthesis remained suppressed. K562 cells were subjected to synchronization prior to and after UV irradiation (24 J/m2) and 18 fractions were collected by centrifugal elutriation. Cell cycle analysis by flow cytometry did not show early apoptotic cells after UV irradiation. The gradual increase in DNA content typical for non-irradiated cells was contrasted by an early S phase block between 2.2 and 2.4 C-values after UV irradiation. Cell cycle dependent chromatin changes after ultraviolet irradiation were seen as a fine fibrillary network covering the mainly fibrous chromatin structures and incompletely folded primitive chromosomes. Based on observations after UV irradiation and on earlier results with cadmium treatment and gamma irradiation, we confirm that typical chromatin changes characteristic to genotoxic agents can be recognized and classified.
Similar content being viewed by others
References
Felsenfeld G (1996) Chromatin unfolds. Cell 86:13–19
Workman JL, Kingston RE (1998) Alteration of nucleosome structure as a mechanism of transcriptional regulation. Annu Rev Biochem 67:545–579
Fordis MC, Anagnou NP, Dean A, Nienhuis AW, Schechter AN (1984) A beta-globin gene, inactive in the K562 leukemic cell, functions normally in a heterologous expression system. Proc Natl Acad Sci USA 81:4485–4489
Kornberg RD (1977) Structure of chromatin. Annu Rev Biochem 46:931–954
Klug A, Butler PJ (1983) The structure of nucleosomes and chromatin. Horiz Biochem Biophys 7:1–41
Kireeva N, Lakonishok M, Kireev I, Hirano T, Belmont AS (2004) Visualization of early chromosome condensation: a hierarchical folding, axial glue model of chromosome structure. J Cell Biol 166:775–785
Nemeth A, Langst G (2004) Chromatin higher order structure: opening up chromatin for transcription. Brief Funct Genomic Proteomic 2:334–343
Banfalvi G, Sooki-Toth A, Sarkar N, Csuzi S, Antoni F (1984) Nascent DNA synthesized reversibly permeable cells of mouse thymocytes. Eur J Biochem 139:553–559
Banfalvi G (1993) Fluorescent analysis of replication and intermediates of chromatin folding in nuclei of mammalian cells. In: Bach PH, Reynolds CH, Clark JM, Mottley J, Poole PL (eds) Biotechnology applications of microinjections, microscopic imaging and fluorescence. Plenum Press, New York, pp 110–119
Banfalvi G, Nagy G, Gacsi M, Roszer T, Basnakian AG (2006) Common pathway of chromosome condensation in mammalian cells. DNA Cell Biol 25:295–301
Banfalvi G (2006) Linear connection of condensing chromosomes in nuclei of synchronized CHO cells. DNA Cell Biol 25:541–545
Banfalvi G, Gacsi M, Nagy G, Kiss ZB, Basnakian AG (2005) Cadmium induced apoptotic changes in chromatin structure and subphases of nuclear growth during the cell cycle in CHO cells. Apoptosis 10:631–642
Nagy G, Gacsi M, Rehak M, Basnakian AG, Klaisz M, Banfalvi G (2004) Gamma irradiation-induced apoptosis in murine pre-B cells prevents the condensation of fibrillar chromatin in early S phase. Apoptosis 9:765–776
Martin SJ, Cotter TG (1991) Ultraviolet B irradiation of human leukemia HL-60 cells in vitro induces apoptosis. Int J Radiat Biol 59:1001–1016
Bogdanov KV, Chukhlovin AB, Zaritskey AI, Frolova OI, Afanasiev BV (1997) Ultraviolet irradiation induces multiple DNA double-strand breaks and apoptosis in normal granulocytes and chronic myeloid leukaemia blasts. Brit J Haematol 98:869–872
Higuchi Y, Mizukami Y, Yoshimoto T (2003) Ultraviolet ray induces chromosomal giant DNA fragmentation followed by internucleosomal DNA fragmentation associated with apoptosis in rat glioma cells. Ann N Y Acad Sci 1010:326–330
Winter DB, Gearhart PJ, Bohr VA (1998) Homogeneous rate of degradation of nuclear DNA during apoptosis. Nucleic Acids Res 26:4422–4425
Lozzio CB, Lozzio BB (1975) Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. Blood 45:321–334
Offer H, Zurer I, Banfalvi G, Rehak M, Falcovitz A, Milyavsky M, Goldfinger N, Rotter V (2001) p53 modulates base excision activity in a cell cycle-specific manner after genotoxic stress. Cancer Res 61:88–96
Basnakian A, Banfalvi G, Sarkar N (1989) Contribution of DNA polymerase delta to DNA replication in permeable CHO cells synchronized in S phase. Nucleic Acids Res 17:4757–4767
Banfalvi G, Poirier L, Mikhailova M, Chou MW (1997) Relationship of repair and replicative DNA synthesis to cell cycle in Chinese hamster ovary (CHO-K1) cells. DNA Cell Biol 16:1155–1160
Castellot JJ Jr, Miller MR, Pardee AB (1978) Animal cells reversibly permeable to small molecules. Proc Natl Acad Sci USA 75:351–355
Moses RE, Richardson CC (1970) Replication and repair of DNA in cells of Escherichia coli treated with toluene. Proc Natl Acad Sci USA 67:674–681
Cleaver JE (1982) Normal reconstruction of DNA supercoiling and chromatin structure in Cockayne syndrome cells during repair of damage from ultraviolet light. Am J Hum Genet 34:566–575
Klein E, Ben-Bassat H, Neumann H, Ralph P, Zeuthen I, Polliack A, Vanky F (1976) Properties of the K562 cell line, derived from a patient with chronic myeloid leukemia. Int J Cancer 18:421–431
Edenberg HJ (1976) Inhibition of DNA replication by ultraviolet light. Biophys J 16:849–860
Banfalvi G, Ujvarosi K, Trencsenyi G, Somogyi C, Nagy G, Basnakaian AG (2007) Cell culture density dependent toxicity and chromatin changes upon cadmium treatment in murine pre-B-cells. Apoptosis 12:1219–1228
Gujuluva CN, Baek JH, Shin KH, Cherrick HM, Park NH (1994) Effect of UV-irradiation on cell cycle, viability and the expression of p53, gadd153 and gadd45 genes in normal and HPV-immortalized human oral keratinocytes. Oncogene 9:1819–1827
Ceruti JM, Scassa ME, Flo JM, Varone CL, Canepa ET (2005) Induction of p19INK4d in response to ultraviolet light improves DNA repair and confers resistance to apoptosis in neuroblastoma cells. Oncogene 24:4065–4080
Acknowledgement
This work was supported by the OTKA grant T042762 (G.B.).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Ujvarosi, K., Hunyadi, J., Nagy, G. et al. Preapoptotic chromatin changes induced by ultraviolet B irradiation in human erythroleukemia K562 cells. Apoptosis 12, 2089–2099 (2007). https://doi.org/10.1007/s10495-007-0118-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10495-007-0118-0