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Topography of genetic loci in the nuclei of cells of colorectal carcinoma and adjacent tissue of colonic epithelium

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Abstract

To determine the influence of increased gene expression and amplification in colorectal carcinoma on chromatin structure, the nuclear distances between pairs of bacterial artificial chromosome (BAC) clones with genomic separation from 800 to 29,000 kb were measured and compared between the tumor and parallel epithelial cells of six patients. The nuclear distances were measured between the loci in chromosomal bands 7p22.3–7p21.3; 7q35–7q36.3; 11p15.5–11p15.4; 20p13; 20p12.2; 20q11.21 and 20q12 where increased expression had been found in all types of colorectal carcinoma. The loci were visualized by three-dimensional fluorescence in situ hybridization using 22 BAC clones. Our results show that for short genomic separations, mean nuclear distance increases linearly with increased genomic separation. The results for some pairs of loci fell outside this linear slope, indicating the existence of different levels of chromatin folding. For the same genomic separations the nuclear distances were frequently shorter for tumor as compared with epithelial cells. Above the initial growing phase of the nuclear distances, a plateau phase was observed in both cell types where the increase in genomic separation was not accompanied by an increase in nuclear distance. The ratio of the mean nuclear distances between the corresponding loci in tumor and epithelium cells decreases with increasing amplification of loci. Our results further show that the large-scale chromatin folding might differ for specific regions of chromosomes and that it is basically preserved in tumor cells in spite of the amplification of many loci.

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Acknowledgements

The authors thank M. Rocchi for the generous supply of BAC clones. The work was supported by the Ministry of Health of the Czech Republic (NC6987-3), Ministry of Education (ME565), the Academy of Sciences of the Czech Republic (A1065203, A5004306) and the Grant Agency of the Czech Republic (GA202/02/0804, GA202/01/0197).

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Authors and Affiliations

  1. Institute of Biophysics, Czech Academy of Sciences, Královopolská 135, 612 65 Brno, Czech Republic

    Emilie Lukášová, Stanislav Kozubek & Martin Falk

  2. Faculty of Informatics, Masaryk University, Botanická 68a, 602 00 Brno, Czech Republic

    Michal Kozubek

  3. Masaryk Memorial Cancer Institute, Žlutý kopec 7, 656 53 Brno, Czech Republic

    Jan Žaloudík, Václav Vagunda & Zdeněk Pavlovský

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  1. Emilie Lukášová
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  2. Stanislav Kozubek
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  3. Martin Falk
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  4. Michal Kozubek
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  5. Jan Žaloudík
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  7. Zdeněk Pavlovský
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Correspondence to Stanislav Kozubek.

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Communicated by T. Hassold

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Lukášová, E., Kozubek, S., Falk, M. et al. Topography of genetic loci in the nuclei of cells of colorectal carcinoma and adjacent tissue of colonic epithelium. Chromosoma 112, 221–230 (2004). https://doi.org/10.1007/s00412-003-0263-3

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  • DOI: https://doi.org/10.1007/s00412-003-0263-3

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