Topography of Genetic Loci in Tissue Samples: Towards New Diagnostic Tool Using Interphase FISH and High-Resolution Image Analysis Techniques

Using single and dual colour fluorescence in situ hybridisation (FISH) combined with image analysis techniques the topographic characteristics of genes and centromeres in nuclei of human colon tissue cells were investigated. The distributions of distances from the centre‐of‐nucleus to genes (centrom...

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Main Authors: I. Koutná, S. Kozubek, J. Žaloudík, M. Kozubek, E. Lukášová, P. A. Matula, E. Bártová, M. Skalníková, A. Cafourková, P. Jirsová
Format: Article
Language:English
Published: Wiley 2000-01-01
Series:Analytical Cellular Pathology
Online Access:http://dx.doi.org/10.1155/2000/369359
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author I. Koutná
S. Kozubek
J. Žaloudík
M. Kozubek
E. Lukášová
P. A. Matula
E. Bártová
M. Skalníková
A. Cafourková
P. Jirsová
author_facet I. Koutná
S. Kozubek
J. Žaloudík
M. Kozubek
E. Lukášová
P. A. Matula
E. Bártová
M. Skalníková
A. Cafourková
P. Jirsová
author_sort I. Koutná
collection DOAJ
description Using single and dual colour fluorescence in situ hybridisation (FISH) combined with image analysis techniques the topographic characteristics of genes and centromeres in nuclei of human colon tissue cells were investigated. The distributions of distances from the centre‐of‐nucleus to genes (centromeres) and from genes to genes (centromeres to centromeres) were studied in normal colon tissue cells found in the neighbourhood of tumour samples, in tumour cell line HT‐29 and in promyelocytic HL‐60 cell line for comparison. Our results show that the topography of genetic loci determined in 3D‐fixed cell tissue corresponds to that obtained for 2D‐fixed cells separated from the tissue. The distributions of the centre‐of‐nucleus to gene (centromere) distances and gene to gene (centromere to centromere) distances and their average values are different for various genetic loci but similar for normal colon tissue cells, HT‐29 colon tumour cell line and HL‐60 promyelocytic cell line. It suggests that the arrangement of genetic loci in cell nucleus is conserved in different types of human cells. The investigations of trisomic loci in HT‐29 cells revealed that the location of the third genetic element is not different from the location of two homologues in diploid cells. We have shown that the topographic parameters used in our experiments for different genetic elements are not tissue or tumour specific. In order to validate high‐resolution cytometry for oncology, further investigations should include more precise parameters reflecting the state of chromatin in the neighbourhood of critical oncogenes or tumour suppresser genes.
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spelling doaj-art-48dc6ed4a8984b2a8752f06f5e1a1edc2025-02-03T01:01:04ZengWileyAnalytical Cellular Pathology0921-89121878-36512000-01-0120417318510.1155/2000/369359Topography of Genetic Loci in Tissue Samples: Towards New Diagnostic Tool Using Interphase FISH and High-Resolution Image Analysis TechniquesI. Koutná0S. Kozubek1J. Žaloudík2M. Kozubek3E. Lukášová4P. A. Matula5E. Bártová6M. Skalníková7A. Cafourková8P. Jirsová9Faculty of Informatics, Masaryk University, Botanická 68a, 602 00 Brno, Czech RepublicInstitute of Biophysics, Academy of Sciences, Královopolská 135, 61265 Brno, Czech RepublicFaculty Hospital of Masaryk University, Bohunice, 635 00 Brno, Czech RepublicFaculty of Informatics, Masaryk University, Botanická 68a, 602 00 Brno, Czech RepublicInstitute of Biophysics, Academy of Sciences, Královopolská 135, 61265 Brno, Czech RepublicFaculty of Informatics, Masaryk University, Botanická 68a, 602 00 Brno, Czech RepublicInstitute of Biophysics, Academy of Sciences, Královopolská 135, 61265 Brno, Czech RepublicFaculty of Informatics, Masaryk University, Botanická 68a, 602 00 Brno, Czech RepublicInstitute of Biophysics, Academy of Sciences, Královopolská 135, 61265 Brno, Czech RepublicInstitute of Biophysics, Academy of Sciences, Královopolská 135, 61265 Brno, Czech RepublicUsing single and dual colour fluorescence in situ hybridisation (FISH) combined with image analysis techniques the topographic characteristics of genes and centromeres in nuclei of human colon tissue cells were investigated. The distributions of distances from the centre‐of‐nucleus to genes (centromeres) and from genes to genes (centromeres to centromeres) were studied in normal colon tissue cells found in the neighbourhood of tumour samples, in tumour cell line HT‐29 and in promyelocytic HL‐60 cell line for comparison. Our results show that the topography of genetic loci determined in 3D‐fixed cell tissue corresponds to that obtained for 2D‐fixed cells separated from the tissue. The distributions of the centre‐of‐nucleus to gene (centromere) distances and gene to gene (centromere to centromere) distances and their average values are different for various genetic loci but similar for normal colon tissue cells, HT‐29 colon tumour cell line and HL‐60 promyelocytic cell line. It suggests that the arrangement of genetic loci in cell nucleus is conserved in different types of human cells. The investigations of trisomic loci in HT‐29 cells revealed that the location of the third genetic element is not different from the location of two homologues in diploid cells. We have shown that the topographic parameters used in our experiments for different genetic elements are not tissue or tumour specific. In order to validate high‐resolution cytometry for oncology, further investigations should include more precise parameters reflecting the state of chromatin in the neighbourhood of critical oncogenes or tumour suppresser genes.http://dx.doi.org/10.1155/2000/369359
spellingShingle I. Koutná
S. Kozubek
J. Žaloudík
M. Kozubek
E. Lukášová
P. A. Matula
E. Bártová
M. Skalníková
A. Cafourková
P. Jirsová
Topography of Genetic Loci in Tissue Samples: Towards New Diagnostic Tool Using Interphase FISH and High-Resolution Image Analysis Techniques
Analytical Cellular Pathology
title Topography of Genetic Loci in Tissue Samples: Towards New Diagnostic Tool Using Interphase FISH and High-Resolution Image Analysis Techniques
title_full Topography of Genetic Loci in Tissue Samples: Towards New Diagnostic Tool Using Interphase FISH and High-Resolution Image Analysis Techniques
title_fullStr Topography of Genetic Loci in Tissue Samples: Towards New Diagnostic Tool Using Interphase FISH and High-Resolution Image Analysis Techniques
title_full_unstemmed Topography of Genetic Loci in Tissue Samples: Towards New Diagnostic Tool Using Interphase FISH and High-Resolution Image Analysis Techniques
title_short Topography of Genetic Loci in Tissue Samples: Towards New Diagnostic Tool Using Interphase FISH and High-Resolution Image Analysis Techniques
title_sort topography of genetic loci in tissue samples towards new diagnostic tool using interphase fish and high resolution image analysis techniques
url http://dx.doi.org/10.1155/2000/369359
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