Koutná I, Kozubek S, Zaloudík J, Kozubek M, Lukásová E, Matula P, Bártová E, Skalníková M, Cafourková A, Jirsová P
Faculty of Informatics, Masaryk University, Brno, Czech Republic.
Anal Cell Pathol. 2000;20(4):173-85. doi: 10.1155/2000/369359.
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.
运用单双色荧光原位杂交（FISH）并结合图像分析技术，对人结肠组织细胞核内基因和着丝粒的拓扑特征进行了研究。在肿瘤样本附近发现的正常结肠组织细胞、肿瘤细胞系HT - 29以及早幼粒细胞HL - 60细胞系中，研究了从细胞核中心到基因（着丝粒）的距离以及从基因到基因（着丝粒到着丝粒）的距离分布，以作比较。我们的结果表明，在三维固定细胞组织中确定的基因座拓扑结构与从组织中分离出的二维固定细胞所获得的拓扑结构相对应。不同基因座的细胞核中心到基因（着丝粒）的距离分布以及基因到基因（着丝粒到着丝粒）的距离分布及其平均值各不相同，但正常结肠组织细胞、HT - 29结肠肿瘤细胞系和HL - 60早幼粒细胞系的情况相似。这表明在不同类型的人类细胞中，细胞核内基因座的排列是保守的。对HT - 29细胞中三体基因座的研究表明，第三个遗传元件的位置与二倍体细胞中两个同源物的位置并无差异。我们已经表明，我们实验中用于不同遗传元件的拓扑参数并非组织或肿瘤特异性的。为了验证用于肿瘤学的高分辨率细胞术，进一步的研究应包括更精确的参数，以反映关键癌基因或肿瘤抑制基因附近染色质的状态。
