Analysis of aclarubicin-induced cell death in human fibroblasts

Authors

  • K. Kania,

    1. Department of Thermobiology, Institute of Biophysics, University of Łódź, Banacha 12/16 Street, 90-237 Łódź, Poland
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    • Centre for Medical Biology of Polish Academy of Sciences, Laboratory Transcriptional Regulation, 106 Lodowa Street, 93-232 Łódź, Poland.

  • K. Matławska-Wąsowska,

    1. Department of Cytogenetics and Plant Molecular Biology, University of Łódź, Banacha 12/16 Street, 90-237 Łódź, Poland
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  • R. Osiecka,

    1. Department of Cytogenetics and Plant Molecular Biology, University of Łódź, Banacha 12/16 Street, 90-237 Łódź, Poland
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  • Z. Jóźwiak

    Corresponding author
    1. Department of Thermobiology, Institute of Biophysics, University of Łódź, Banacha 12/16 Street, 90-237 Łódź, Poland
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Corresponding author. Tel.: +48 042 6354479. zjozwiak@biol.uni.lodz.pl

Abstract

In the present study we investigated the mode of cell death induced by aclarubicin (ACL) in trisomic (BB) and normal (S-2) human fibroblasts. Cells were incubated with ACL for 2 h and then cultured in drug-free medium for up to 96 h. Using fluorescence microscopy, agarose gel electrophoresis and comet assay we demonstrate that ACL induced time-dependent morphological and biochemical changes in both cell types. The population of apoptotic cells, analysed by acridine orange and ethidium bromide nuclear staining reached its maximum at 24–48 h. Prolonged post-treatment time progressively increased the level of necrotic cells. At 24–48 h time points we also observed a significant increase in caspase-3 activity, oligonucleosomal DNA fragmentation and DNA strand breaks. Cotreatment of cells with the specific caspase-3 inhibitor Ac-DEVD-CHO partly reduced the extent of apoptosis and necrosis and DNA degradation. In conclusion, trisomic and normal fibroblasts demonstrate similar response to aclarubicin treatment. Drug induced the apoptotic and necrotic pathway of cell death that was mediated by caspase-3.

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