Targeting multiple genetic aberrations in isolated tumor cells by spectral fluorescence in situ hybridization

Slovak, Marilyn L.; Zhang, Feiyu; Tcheurekdjian, Lucene; Bobadilla, Dolores; Bedell, Victoria; Arber, Daniel A.; Persons, Diane L.; Sosman, Jeffrey A.; Murata-Collins, Joyce

doi:10.1016/s0361-090x(02)00063-6

Cited by 8 publications

(3 citation statements)

References 13 publications

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“…Cytogenetic methods have thus far mainly been applied to cultured cells, such as lymphocytes. Although it has recently become possible to apply cytogenetic methods, such as FISH, on interphase cells (Slovak et al 2001), no results are available for mouse organs and tissues to directly compare with our results. Indeed, the difficulty in analyzing genome rearrangements with cytogenetic methods or methods based on endogenous reporter genes was the reason to generate the lacZ plasmid model system in the first place (Gossen and Vijg 1993).…”

Section: ‫4מ‬mentioning

confidence: 99%

Genome Dynamics in Aging Mice

Dollé¹,

Vijg²

2002

Genome Res.

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Random spontaneous genome rearrangements are difficult to detect in vivo, especially in postmitotic tissues. Using a lacZ-plasmid reporter mouse model, we have previously presented evidence for the accumulation of large genome rearrangements in various tissues, including postmitotic tissues, during aging. These rearrangements, which were found to be organ-specific and to increase with age, have one breakpoint in the lacZ-reporter locus and the second elsewhere in the mouse genome. In this present work, we have used a mouse genome sequence database to physically characterize a total of 49 genome rearrangements in the brain, heart, and liver from young and old mice at two lacZ-plasmid reporter loci. Half of all breakpoints in the mouse genome occurred in chromosomes 3 and 4, each carrying a lacZ-reporter cluster, at distances varying from <100 kb to 66 Mb, indicating intrachromosomal deletions or inversions. The other half of the breakpoints in the mouse genome was found randomly on any of the other chromosomes, indicating translocations. Alternatively, part of the intra-and extrachromosomal events could involve transpositions. Regions of extended homology were not found at the breakpoints. These results lead us to postulate potential mechanisms for the origin of large genome rearrangements in mouse tissues and to predict their possible impact as a potential cause of aging.

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Section: ‫4מ‬mentioning

confidence: 99%

Genome Dynamics in Aging Mice

Dollé¹,

Vijg²

2002

Genome Res.

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“…This procedure allows for evaluation of 500 to 2000 mitotic cells [20•,21] and may be used in combination with immunophenotyping of flow-sorted cell populations [22][23][24] or simultaneously with multiple DNA probes. The last process increases the sensitivity of the FISH assay to 10 -3 to 10 -4 [25]. The main technical limitations of FISH are false-negative results caused by poor hybridization and false-positive results caused by nonspecific probe hybridization or chance co-localization of signals in interphase nuclei mimicking a translocation event.…”

Section: Cytogenetic Evaluationmentioning

confidence: 99%

Monitoring bcr-abl by polymerase chain reaction in the treatment of chronic myeloid leukemia

Radich

2003

Curr Oncol Rep

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The elucidation of the molecular biology of chronic myeloid leukemia (CML) has provided a paradigm for understanding leukemogenesis, targeted drug development, and disease monitoring at the molecular level. Minimal residual disease (MRD) monitoring by fluorescence in situ hybridization and polymerase chain reaction (PCR) has become an important tool in predicting relapse after allogeneic transplant, allowing for early intervention strategies such as donor lymphocyte infusion. MRD monitoring is important for assessment of disease status in patients who obtain a complete cytogenetic remission, and this approach is likely to play an important role in following patients to determine who will relapse on imatinib mesylate therapy. This review focuses primarily on MRD monitoring by PCR.

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“…Die Interphasezytogenetik bietet auch den groûen Vorteil, neben chromosomalen DNA-Sonden auch prognostisch wichtige Gen-Sonden simultan zu verwenden. Durch einen solchen Ansatz könnten gleichzeitig mehrere eingesetzte Sonden eine umfassende Charakterisierung zirkulierender Tumorzellen ermöglichen[18]. Die Signifikanz für das klinische Management onkologischer Patienten wäre enorm.Weiterhin demonstrieren unsere Daten, daû selbst bei frühen Tumorstadien des Mammakarzinoms, wie T1N0, ein signifikanter Anteil der Patientinnen zirkulierende Tumorzellen aufweist.…”

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Interphasezytogenetik mit DNA-Sonden für Chromosom 8 zur Detektion zirkulierender Tumorzellen beim Mammakarzinom

Ghadimi¹,

Uhr²,

Tucker³

et al. 2001

Zentralbl Chir

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The detection of micrometastases in the bone marrow or peripheral blood of cancer patients is increasingly used for a more sensitive tumor staging and prognostication. The potential value of the currently used techniques for the detection of epithelial antigens by RT-PCR or immunohistochemistry in respect of specificity is currently controversially discussed. In the present study we demonstrate a new approach which enables the direct visualization of the tumor specific alteration of chromosome 8 in circulating tumor cells. We have therefore studied breast cancer patients with various tumor stages and tried to determine the frequency of circulating tumor cells in the peripheral blood by using interphase cytogenetics for chromosome 7 and 8. Imprints of primary breast cancers and cytospins with circulating tumor cells of corresponding patients were studied in a blinded fashion. The blood samples were generated by immunomagnetic enrichment of circulating tumor cells from peripheral blood by ferrofluid and centrifugation onto cover slips. These cytospins were then hybridized with centromer probes 7 and 8. After analyzing 27 patients with benign as well as malignant breast tumors we can demonstrate that the chromosomal pattern between malignant tumor and corresponding circulating tumor cells is identical. Furthermore, the detection of circulating tumor cells directly correlates with the primary tumor stage. We did not find any cells with chromosome 8 alterations in the patients with benign disease. Surprisingly, even in early breast cancers (T1N0) interphase cytogenetics identified circulating tumor cells in 2 out of 4 patients. In conclusion, interphase cytogenetics represent a non-invasive, sensitive and specific assay for the direct visualization of circulating tumor cells in the peripheral blood. The prognostic value of these findings remains to be further evaluated in larger prospective studies.

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Targeting multiple genetic aberrations in isolated tumor cells by spectral fluorescence in situ hybridization

Cited by 8 publications

References 13 publications

Genome Dynamics in Aging Mice

Genome Dynamics in Aging Mice

Monitoring bcr-abl by polymerase chain reaction in the treatment of chronic myeloid leukemia

Interphasezytogenetik mit DNA-Sonden für Chromosom 8 zur Detektion zirkulierender Tumorzellen beim Mammakarzinom

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