Detection of chromosome aberrations in metaphase and interphase tumor cells by in situ hybridization using chromosome-specific library probes

Abstract

Summary. Chromosome aberrations in two glioma cell lines were analyzed using biotinylated DNA library probes that specifically decorate chromosomes 1, 4, 7, 18 and 22 from pter to qter. Numerical changes, deletions and rearrangements of these chromosomes were readily visualized in metaphase spreads, as well as in early prophase and interphase nuclei. Complete chromosomes, deleted chromosomes and segments of translocated chromosomes were rapidly delineated in very complex karyotypes. Simultaneous hybridizations with additional subregional probes were used to fllrther define aberrant chromosomes. Digital image analysis was used to quantitate the total complement of specific chromosomal DNAs in individual metaphase and interphase cells of each cell line. In spite of the fact that both glioma lines have been passaged in vitro for many years, an under-representation of chromosome 22 and an over-representation of chromosome 7 (specifically 7p) were observed. These observations agree with previous studies on gliomas. In addition, sequences of chromosome 4 were also found to be under-represented, especially in TC 593. These analyses indicate the power of these methods for pinpointing chromosome segments that are altered in specific types of tumors.

Frequent loss of chromosome 14 in atypical and malignant meningioma: identification of a putative `tumor progression' locus

“…All probes were labeled with biotin by nick translation and suppression hybridization was performed as described Cremer et al, 1988), with the following exceptions. Each hybridization reaction (10 ml volume) contained 300 ng of probe, 2.0 mg of human Cot-1 DNA (GIBCO BRL, Gaithersburg, MD) and 8.0 mg of salmon sperm DNA in a cocktail of 50% formamide, 10% Dextran Sulfate and 66SSC (pH 7.0).…”

Cytometry and genetics

“…REVERSE PAINTING Flow-sorted chromosome-specific DNA libraries constructed by the National Laboratory Gene Library Project found an arguably wider ranging use in the form of chromosome paints (19,(25)(26)(27). The libraries, fluorescently labeled and hybridized to metaphase spreads, allowed the whole chromosome corresponding to the original sorted chromosome to be directly visualized.…”

Painting of human chromosomes with probes generated from hybrid cell lines by PCR with Alu and L1 primers

“…Metaphase chromosome spreads were obtained from hybrid cell lines and from phytohemagglutinin (PHA)-stimulated normal male and female human blood lymphocytes using standard procedures (Cremer et al 1988). Preparations were stored in 70% ethanol at 4~ until use.…”

“…Recently, chromosomal in situ suppression (CISS-) hybridization protocols have been developed that allow the use of complex probe sets, such as complete DNA libraries derived from sorted human chromosomes, to visualize individual chromosomes in a highly specific manner (Pinkel et al 1988: Lichter et al 1988a: Cremer et al 1988. This technique, which is also referred to as chromosome painting (Pinkel et al, 1988), has provided a novel approach to improve the cytogenetic evaluation of hybrid cell lines (Kievits et al 1990: Boyle et al 19901.…”