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Colorectal Cancer pp 35–50Cite as

Fluorescence In Situ Hybridization

Application in Cancer Research and Clinical Diagnostics

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Part of the book series: Methods in Molecular Medicine ((MIMM,volume 50))

Abstract

An opportunity to look inside of the individual cell for the direct visualization in situ of “what happened?” is the most wonderful feature offered by fluorescence in situ hybridization (FISH). DNA in situ hybridization is a technique that allows the visualization of defined sequences of nucleic acids within the individual cells. The method is based on the site specific annealing (hybridization) of single-stranded labeled DNA fragments (probes) to denatured, homologous sequences (targets) on cytological preparations, like metaphase chromosomes, interphase nuclei, or naked chromatin fibers. Visualization of hybridization sites becomes possible after detection steps by using a wide spectrum of the fluorescent dyes available.

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References

  1. Pack, S. D., Tanigami, A., Sato, T., Ledbetter, D., and Fukuda, M. N. (1997) Assignment of trophoblast/endometrial epithelium cell adhesion molecule trophinin gene Tro to human chromosome bands Xp11. 21-11. 22 by in situhybridization Cytogenet. Cell Genet. 79 (1-2), 123–124.

    Article  Google Scholar 

  2. Pack, S. D., Pak, E., Tanigami, A., Ledbetter, D. H., and Fukuda, M. N. (1999) Assignment of bystin gene BYSL to human chromosome band 6p21. 1 by in situhybridization Cytogenet. Cell Genet. 83, 76–77

    Article  Google Scholar 

  3. Nederlof, P., van der Flier, S., Raap, A. K., Tanke, H. J., van der Ploeg, M., Kornips, F., and Geraedts, J. P. M. (1989) Detection of chromosome aberrations in interphase tumor nuclei by non-radioactive in situ hybridization. Cancer Genet. Cytogenet. 42, 87–98.

    Article  CAS  PubMed  Google Scholar 

  4. Tkachuk, D. C., Pinkel, D., Kuo, W.-L., Weier, H.-U., and Gray, J. W. (1991) Clinical applications of fluorescence in situ hybridization. Genet. Anal. Techn. Appl. 8, 67–74.

    CAS  Google Scholar 

  5. Ried, T., Lengauer, C., Cremer, T., Wiegant, J., Raap, A. K., van der Ploeg, M., Groitl, P., and Lipp, M. (1992) Specific metaphase and interphase detection of the breakpoint region in 8q24 of Burkitt lymphoma cells by triple-color fluorescence in situ hybridization. Genes Chrom. Cancer 4, 69–74.

    Article  CAS  PubMed  Google Scholar 

  6. Lengauer, C., Green, E. D., and Cremer, T. (1992) Fluorescence in situ hybridization of YAC clones after Alu-PCR amplification. Genomics 13, 826–828.

    Article  CAS  PubMed  Google Scholar 

  7. Ried, T., Lengauer, C., Lipp, M., Fischer, C., Cremer, T., and Ward, D. C. (1993) Evaluation of the utility of interphase cytogenetics to detect residual cells with a malignant genotype in mixed cell populations: a Burkitt lymphoma model. DNA CellBiol. 12, 637–643.

    Article  CAS  Google Scholar 

  8. Demetrick, D. J. (1996) The use of archival frozen tumor tissue imprint specimens for fluorescence in situ hybridization. Modern Pathol. 9 (2), 133–136.

    Google Scholar 

  9. Pack, S., Vortmeyer, A., Pak, E., Liotta, L., and Zhuang, Z. (1997) Detection of single metastatic cells in lymph node tissue by fluorescent in-situ hybridization. Lancet 350, 264–265.

    Article  CAS  PubMed  Google Scholar 

  10. Pack, S. D., Turner, M. L., Zhuang, Z., Vortmeyer, A. O., Boni, R, Scarulis, M., Marx, S. J., and Darling, T. N. (1998) Cutanious tumors in patients with multiple endocrine neoplasia type I show allelic deletion of the MEN1 gene J. Invest. Dermatol. 110, 438–441.

    Article  CAS  PubMed  Google Scholar 

  11. Pack, S., Boni, R., Vortmeyer, A., Pak, E., and Zhuang, Z. (1998) Detection of gene deletion in single metastatic tumor cells in the exicion margin of a primary cutanious melanoma by fluorescent in situ hybridization. J. Natl. Cancer Inst. 90(10), 782–783.

    Google Scholar 

  12. Kuwano, A., Ledbetter, S. A., Dobins, W. B., Emanuel, B. S., and Ledbetter, D. H. (1991) Detection of deletions and cryptic translocations in Miller-Dieker syndrome by in situ hybridization. Am. J. Hum. Genet. 49(4), 707–714.

    CAS  PubMed  Google Scholar 

  13. Hirota, H., Matsuoka, R., Kimura, M., Imamura, S., Joho, K., Ando, M., Takao, A., and Momma, K. (1996) Molecular cytogenetic diagnosis of Williams syndrome. Am. J. Med. Genet. 64(3), 473–477.

    Article  Google Scholar 

  14. Shaffer, L. G., Kennedy, G. M., Spikes, A. S., and Lupski J. R. (1997) Diagnosis of CMT1A duplications and HNPP deletions by interphase FISH: implications for testing in the cytogenetics laboratory. Am. J. Med. Genet. 69(3), 325–331.

    Article  Google Scholar 

  15. Krantz, I. D., Rand, E. B., Genin, A., Hunt, P., Jones, M., Louis, A. A., et al. (1997) Deletions of 20p12 in Allagille syndrome: frequency and molecular characterization. Am. J. Med. Genet. 70(1), 80–86.

    Article  Google Scholar 

  16. Kumar, S., Pack, S. D, Kumar, D., Walker, R., Quezado, M., Zhuang, Z., et al. (1999) Detection of EWS-FLI-1 fusion in Ewing′s sarcoma/peripheral primitive neuroectodermal tumor by fluorescence in-situ hybridization using formalin-fixed paraffin-embedded tissue. Human Pathol. 30(3), 324–330.

    Article  Google Scholar 

  17. Chan, C. C., Pack, S. D., Pak, E., Tsokos, M., and Zhuang, Z. (1999) Translocation of chromosomes 11 and 22 in choroidal metastatic Ewing′ s Sarcoma detected by fluorescence in-situ hybridization. Am. J. Opthalmol. 127(2), 226–228.

    Article  Google Scholar 

  18. Pack, S. D., Zbar, B., Pak, E., Ault, D. O., Humphrey, J., Pham, T., et al. (1999) Germline deletions in VHL syndrome detected by fluorescence in situ hybridization (FISH). Cancer Res. 59(2), 5560–5563.

    Google Scholar 

  19. Stilgenbauer, S., Döhner, H., Bulgary-Mörschel, M., Weitz, S., Bentz, M., and Lichter, P. (1993) Retinoblastoma gene deletion in chronic lymphoid leukemias: a combined metaphase and interphase cytogenetic study. Blood 81, 2118–2124.

    CAS  PubMed  Google Scholar 

  20. Vortmeyer, A., Choo, D., Pack, S., Oldfield, E., and Zhuang, Z. (1997, July 2) VHL gene alterations associated with endolymphatic sac tumors. J. Natl. Cancer Inst. 89(13), 970–972.

    Google Scholar 

  21. Zhuang, Z., Vortmeyer, A. O., Pack, S., Huang, S., Pham, T., Wang, C., et al. (1997) Somatic mutations of the MEN1 tumor suppressor gene in sporadic gastrinomas and insulinomas. Cancer Res. 57(21), 4682–4686.

    Google Scholar 

  22. Zhuang, Z., Ezzat, S., Vortmeyer, A. O., Weil, R., Oldfield, E. H., et al. (1997) Mutations of the MEN1 tumor suppressor gene in pituitary tumors. Cancer Res. 57(24), 5446–5451.

    Google Scholar 

  23. Lubensky, I. A., Pack, S., Ault, D., Vortmeyer, A. O., Libutti, S. K., Choyke, P. L., et al. (1998) Multiple neuroendocrine tumors of the pancreas in von Hippel-Lindau Disease Patients: Histopathologic and Molecular Genetic Analysis. Am. J. Pathol. 153(1), 1378–1384.

    Google Scholar 

  24. Zhuang, Z., Park, W-S., Pack, S. D., Schmidt, L., Vortmeyer, A. O., Pak, E., et al. (1998) Trisomy 7 harboring non-random duplication of the mutant c-met proto-oncogene allele in hereditary papillary renal carcinomas. Nat. Genet. 20, 66–69.

    Article  CAS  PubMed  Google Scholar 

  25. Thibleton, C., Pack, S., Sakai, A., Beaty, M., Pak, E., Vortmeyer, A. O., et al. (1999) Allelic loss of 11q13 as detected by MEN1-FISH is not associated with mutation of the MEN1 gene in lymphoid neoplasms Leukemia 13(1), 85–91.

    Google Scholar 

  26. Kallioniemi, A., Kallioniemi, O.-P, Sudar, D., Rutovitz, D., Gray, J. W., Waldman, F., and Pinkel, D. (1992) Comparative genomic hybrization for molecular cytogenetic analysis of solid tumors. Science 258, 818–821.

    Article  CAS  PubMed  Google Scholar 

  27. Pack, S. D., Karkera, J. D., Zhuang, Z., Pak, E., Hwu, P., Balan, K. W., et al. (1999) Molecular cytogenetic fingerprinting of esophageal squamous cell carcinoma by comparative genomic hybridization (CGH) reveals consistent pattern of chromosomal alterations. Genes Chrom. Cancer 25, 160–168.

    Article  CAS  PubMed  Google Scholar 

  28. Schrock, E., du Manoir, S., Veldman, T., Schoell, B., Wienberg, J., Ferguson-Smith, M. A., et al. (1996) Multicolor spectral karyotyping of human chromosomes. Science 273(5274), 494–497.

    Article  CAS  PubMed  Google Scholar 

  29. Cremer, T., Lichter, P., Borden, J., Ward, D. C., and Manuelidis, L. (1988) Detection of chromosome aberrations in metaphase and interphase tumor cells by in situ hybridization using chromosome specific library probes. Hum. Genet. 80, 235–246.

    Article  CAS  PubMed  Google Scholar 

  30. Ploem, J. S. and Tanke, H. J. (1987) Introduction to fluorescence microscopy, in RMS microscopy handbooks series, No. 10, Oxford Science Publications.

    Google Scholar 

  31. Marcus, D. A. (1988) High-performance optical filters for fluorescence analysis. Cell Motil. Cytoskelet. 10, 62–70.

    Article  CAS  Google Scholar 

  32. Johnson, C. V., McNeil, J. A., Carter, K. C., and Lawrence, J. B. (1991) A simple, rapid technique for precise mapping of multiple sequences in two colors using a single optical filter set. Genet. Anal. Techn. Appl. 8, 24–35.

    Google Scholar 

  33. Hiraoka, Y., Sedat, J. W., and Agard, D. A. (1987) The use of a charge-coupled device for quantitative optical microscopy of biological structures. Science 238, 36–41.

    Article  CAS  PubMed  Google Scholar 

  34. Cremer, C. and Cremer, T. (1978) Considerations on a laser-scanning microscope with high resolution and depth of field. Microscopia Acta 81, 31–44.

    CAS  Google Scholar 

  35. Haaf, T. and Ward, D. C. (1994) Structural analysis of alpha-satellite DNA and centromere proteins using extended chromatin and chromosomes. Hum. Mol. Gen. 3(5), 697–709.

    Article  Google Scholar 

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Author information

Authors and Affiliations

  1. Laboratory of Pathology, National Cancer Institute, NIH, Bethesda, MD

    Svetlana D. Pack & Zhengping Zhuang

Authors
  1. Svetlana D. Pack
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  2. Zhengping Zhuang
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Editor information

Editors and Affiliations

  1. University of Virginia Health System, Charlottesville, VA

    Steven M. MD

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© 2001 Humana Press Inc.

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Pack, S.D., Zhuang, Z. (2001). Fluorescence In Situ Hybridization. In: M., S. (eds) Colorectal Cancer. Methods in Molecular Medicine, vol 50. Humana Press. https://doi.org/10.1385/1-59259-084-5:35

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  • DOI: https://doi.org/10.1385/1-59259-084-5:35

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-767-0

  • Online ISBN: 978-1-59259-084-1

  • eBook Packages: Springer Protocols

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