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Architecture of chromosomal territories in apoptotic cells

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BÁRTOVÁ Eva FOJTOVÁ Miloslava SOUČEK Karel MATULA Pavel PALŠA Stanislav KOZUBEK Stanislav

Year of publication 2003
Type Article in Proceedings
Conference Biophysics of the Genome
MU Faculty or unit

Faculty of Informatics

Field Genetics and molecular biology
Keywords human genome structure
Description The nuclear architecture of selected chromosomes in apoptotic nuclei of human leukemic cells K-562 and HL-60 was investigated in the presented paper. Etoposide and prolonged confluence were used for the induction of apoptosis. DAPI- as well as TUNEL- labelling of apoptotic nuclear bodies was combined with visualisation of chromosomal territories by the FISH technique. Simultaneous vital staining by Annexin V, PI and Hoechst 33342 was applied in order to distinguish apoptotic, necrotic and intact cell fraction of tested population. Our FISH analyses revealed that the 3D- structure of apoptotic nuclei as well as the 3D- structure of apoptotic bodies is preserved in formaldehyde fixed cells. High molecular weight DNA fragmentation was determined in apoptotic K-562 cells in contrast to oligonucleosomal cleavage observed in apoptotic HL In comparison with the control, an increased number of centromeric FISH signals were observed in prolonged confluence treated K-562 cells induced to apoptosis. This finding can be explained either as a consequence of apoptosis or by polyploidization. Sequential staining of the same apoptotic nuclei by the FISH and TUNEL techniques revealed that chromosomal territory segmentation precedes the formation of nuclear apoptotic bodies.
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