In Situ Detection of Supernumerary Aberrations of Chromosome-Specific Repetitive DNA Targets in Interphase Nuclei in Human Melanoma Cell Lines and Tissue Sections
| Autor: | Dirk J. Ruiter, O. Moesker, Goos N.P. van Muijen, Peter E.J. de Wit, Anton H. N. Hopman, Dominique Smeets, Johan L. M. Beck |
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| Jazyk: | angličtina |
| Předmět: |
Male
Pathology medicine.medical_specialty Transplantation Heterologous Population Mice Nude Aneuploidy In situ hybridization Dermatology Biology Biochemistry Mice Tumor Cells Cultured medicine Animals Humans education Melanoma Molecular Biology Aged Chromosome Aberrations Chromosome 7 (human) education.field_of_study Nucleic Acid Hybridization Karyotype Cell Biology Flow Cytometry medicine.disease Chromosomes Human Pair 1 Tumor progression Karyotyping Cutaneous melanoma Melanocytes Chromosomes Human Pair 7 Neoplasm Transplantation |
| Zdroj: | Journal of Investigative Dermatology. (4):450-458 |
| ISSN: | 0022-202X |
| DOI: | 10.1111/1523-1747.ep12499853 |
| Popis: | The use of non-radioactive in situ hybridization (ISH) with chromosome-specific repetitive DNA probes to study genomic changes, aneuploidy, and heterogeneity during melanocytic tumor progression, relies on its applicability to non-mitotic interphase nuclei, present in cell suspensions and tissue sections. Therefore, we studied the feasibility of detecting numerical aberrations with respect to the (peri-) centromere regions of chromosomes 1 and 7 in intact nuclei of two human melanoma cell lines with different metastatic behavior in nude mice. In addition, we used paraffin sections from xenograft lesions, obtained by inoculation of these cell lines in nude mice (subcutaneous tumors and spontaneous lung metastases). Paraffin sections from the original primary cutaneous melanoma (with a subepidermal and a dermal part) and two loco-regional metastases were also studied, one of which was the source for the cell lines. These cells and tissues represent examples of materials used in different approaches to the study of melanocytic tumor progression. Regarding the targeted sequences, ISH analysis showed that both cell lines were heterogeneous and aneuploid. The results correlated well with those obtained by ISH on metaphase spreads. Differences between the lines, which could not be detected by flow-cytometric or conventional karyotyping analysis, included data suggestive of a polyploid subpopulation and an extra copy of chromosome 7 in the metastasizing cell line. The polyploid population could be detected also in the paraffin sections of the corresponding subcutaneous xenografts and lung metastases in the mice. Both areas in the patients' primary melanoma could be evaluated separately and showed similar supernumerary aberrations of the chromosome-specific targets. These abnormalities matched those found in both metastases. Our results demonstrate that ISH can be used to visualize genomic abnormalitites at the single-cell level in melanocytic nuclei in their natural context, which makes it a promising tool in the histopathology of melanocytic lesions and in the study of melanocytic tumor progression. |
| Databáze: | OpenAIRE |
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