Generation of a Specific Fluorescence In Situ Hybridization Test for the Detection of Ovarian Carcinoma Cells.
Amelie LimburgXueqian QianBernice BrechtefeldNina HedemannInken FlörkemeierChristoph RogmansLeticia Oliveira-FerrerNicolai MaassNorbert ArnoldDirk Olaf BauerschlagJörg Paul WeimerPublished in: Biomedicines (2024)
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.
Keyphrases
- single molecule
- genome wide
- living cells
- stem cells
- bone marrow
- genome wide association study
- endothelial cells
- quantum dots
- energy transfer
- peripheral blood
- genome wide association
- dna methylation
- high throughput
- copy number
- high grade
- circulating tumor
- pet ct
- positron emission tomography
- induced pluripotent stem cells
- circulating tumor cells
- high density