Generation of a Specific Fluorescence In Situ Hybridization Test for the Detection of Ovarian Carcinoma Cells

Autor: Amelie Limburg, Xueqian Qian, Bernice Brechtefeld, Nina Hedemann, Inken Flörkemeier, Christoph Rogmans, Leticia Oliveira-Ferrer, Nicolai Maass, Norbert Arnold, Dirk O. Bauerschlag, Jörg Paul Weimer
Jazyk: angličtina
Rok vydání: 2024
Předmět:
Zdroj: Biomedicines, Vol 12, Iss 6, p 1171 (2024)
Druh dokumentu: article
ISSN: 2227-9059
DOI: 10.3390/biomedicines12061171
Popis: Examinations of ovarian cancer cells require the ability to identify tumor cells. Array-based comparative genome hybridization (aCGH) on 30 ovarian carcinomas (OC) identified three genomic loci (8q24.23; 17p12; 18q22.3) over- or under-represented in OC. A fluorescence in situ hybridization (FISH) probe of these three loci is intended to identify tumor cells by their signal pattern deviating from a diploid pattern. Human DNA from these three loci is isolated from bacterial artificial chromosomes (BAC), amplified and labeled with fluorescent dyes. After a standard FISH procedure, 71 OC suspensions from primary tumors, three OC cell lines, three lymphocyte suspensions, and one mesenchymal cell line LP-3 are analyzed with a fluorescence microscope. On average, 15% of the lymphocytes deviate from the expected diploid signal pattern, giving a cut-off of 36%. If this value is exceeded, tumor cells are detected. The mesenchymal cell line LP-3 shows only 21% as a negative control. The OC cell lines as positive controls exceed this value at 38%, 67%, and 54%. Of the 71 OC primary cultures, four cases fell below this cut-off as false negatives. In the two-sample t-test, the percentages of conspicuous signal patterns differ significantly.
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