Skip to main content

Comparative Genomic Hybridization of Human Lung Cancer

  • Protocol
Lung Cancer

Part of the book series: Methods in Molecular Medicineā„¢ ((MIMM,volume 75))

  • 645 Accesses

Abstract

Lung cancer is a highly aggressive neoplasm, a characteristic that is reflected by the multitude of genetic aberrations detectable on the chromosomal and molecular level. In order to understand these seemingly chaotic chromosomal alterations, we have performed Comparative Genomic Hybridization (CGH) on a large collective of human lung carcinomas. CGH was one of the first screening methods used to analyze tumor genomes for genetic imbalances (1,2), and has rapidly become a popular tool for a comprehensive molecular cytogenetic analysis of solid tumors. The method is based on the hybridization of differentially labeled tumor and reference DNA on normal chromosome metaphase spreads. Both genomes compete for complementary binding sites on the chromosomal DNA. In the case of an amplification in tumor DNA, more DNA will hybridize to the corresponding band or chromosomal arm, whereas deletions will allow the binding of the normal DNA to competitively prevail. Detection of each genome is facilitated by the use of distinct fluorochrome labels. DNA imbalances are determined by comparing the intensity of the fluorescence signals along individual chromosomes. DNA gains are potentially associated with the activation of proto-oncogenes and DNA losses with the inactivation of tumor-suppressor genes.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Kallioniemi, O. P., Kallioniemi, A., Sudar, D., Rutovitz, D., Gray, J. W., Waldman, F., and Pinkel, D. (1992) Comparative genomic hybridization for molecular cytogenetic analysis of solid tumors. Science 258, 818ā€“821.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  2. du Manoir, S., Speicher, M. R., Joos, S., Schrƶck, E., Popp, S., Dƶhner, H., et al. (1993) Detection of complete and partial chromosome gains and losses by comparative genomic in situ hybridization. Human Genet. 90, 590ā€“610.

    ArticleĀ  Google ScholarĀ 

  3. Speicher, M. R., du Manoir, S., Schrƶck, E., Holtgreve-Grez, H., Schoell, B., Lengauer, C., et al. (1993) Molecular cytogenetic analysis of formalin-fixed, paraffin-embedded solid tumors by comparative genomic hybridization after universal DNA-amplification. Human Mol. Genet. 2, 1907ā€“1914.

    ArticleĀ  CASĀ  Google ScholarĀ 

  4. Isola, J., DeVries, S., Chu, L., Ghazvini, S., and Waldman, F. (1994) Analysis of changes in DNA sequence copy number by comparative genomic hybridization in archival paraffin-embedded tumor samples. Am. J. Pathol. 145, 1301ā€“1308.

    PubMedĀ  CASĀ  Google ScholarĀ 

  5. Wiltshire, R. N., Duray, P., Bittner, M. L., Visakorpi, T., Meltzer, P. S., Tuthill, R. J., et al. (1995) Direct visualization of the clonal progression of primary cuatneous melanoma: application of tissue microdissection and comparative genomic hybridization. Cancer Res. 55, 3954ā€“3957.

    PubMedĀ  CASĀ  Google ScholarĀ 

  6. Kuukasjarvi, T., Tanner, M., Pennanen, S., Karhu, R., Visakorpi, T., and Isola, J. (1997) Optimizing DOP-PCR for universal amplification of small DNA samples in comparative genomic hybridization. Genes Chromosomes Cancer 18, 94ā€“101.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  7. Kallioniemi, O. P., Kallioniemi, A., Piper, J., Isola, J., Waldman, F. M., Gray, J. W., and Pinkel, D. (1994) Optimizing comparative genomic hybridization for analysis of DNA sequence copy number changes in solid tumors. Genes Chromosomes Cancer 10, 231ā€“243.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  8. Karhu, R., Kahkonen, M., Kuukasjarvi, T., Pennanen, S., Tirkkonen, M., and Kallioniemi, O. P. (1997) Quality control of CGH: impact of metaphase chromosomes and the dynamic range of hybridization. Cytometry 28, 198ā€“205.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  9. Bentz, M., Plesch, A., Stilgenbauer, S., Dohner, H., and Lichter, P. (1998) Minimal sizes of deletions detected by comparative genomic hybridization. Genes Chromosomes Cancer 21, 172ā€“175.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  10. Schena, M., Shalon, D., Davis, R. W., and Brown, P. O. (1995) Quantitative monitoring of gene expression patterns with a complementary DNA microarray. Science 270, 467ā€“470.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  11. Kraus, J., Weber, R. G., Cremer, M., Seebacher, T., Fischer, C., Schurra, C., et al. (1997) High-resolution comparative hybridization to combed DNA fibers. Hum. Genet. 99, 374ā€“380.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  12. Solinas-Toldo, S., Lampel, S., Stilgenbauer, S., Nickolenko, J., Benner, A., Dohner, H., et al. (1997) Matrix-based comparative genomic hybridization: biochips to screen for genomic imbalances. Genes Chromosomes Cancer 20, 399ā€“407.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  13. Pinkel, D., Segraves, R., Sudar, D., Clark, S., Poole, I., Kowbel, D., et al. (1998) High resolution analysis of DNA copy number variation using comparative genomic hybridization to microarrays. Nat. Genet. 20, 207ā€“211.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  14. Pollack, J. R., Perou, C. M., Alizadeh, A. A., Eisen, M. B., Pergamenschikov, A., Williams, C. F., et al. (1999) Genome-wide analysis of DNA copy-number changes using cDNA microarrays. Nat. Genet. 23, 41ā€“46.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  15. Roth, K., Wolf, G., Dietel, M., and Petersen, I. (1997) Image analysis for Comparative Genomic Hybridization based on a karyotyping program for Windows. Anal. Quant. Cytol. Histol. 19, 461ā€“474.

    PubMedĀ  CASĀ  Google ScholarĀ 

  16. Petersen, I., Bujard, M., Petersen, S., Wolf, G., Goeze, A., Schwendel, A., et al. (1997) Patterns of chromosomal imbalances in adenocarcinoma and squamous cell carcinoma of the lung. Cancer Res. 57, 2331ā€“2335.

    PubMedĀ  CASĀ  Google ScholarĀ 

  17. Petersen, I., Langreck, H., Wolf, G., Schwendel, A., Psille, R., Vogt, P., et al. (1997) Small cell lung cancer is characterized by a high incidence of deletions on chromosomes 3p, 4q, 5q, 10q, 13q and 17p. Br. J. Cancer 75, 79ā€“86.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  18. Schwendel, A., Langreck, H., Reichel, M. B., Schrƶck, E., Ried, T., Dietel, M., and Petersen, I. (1997) Comparative Genomic Hybridization reveals a common pattern of genetic changes in primary small cell lung carcinomas and their metastases. Int. J. Cancer 74, 86ā€“93.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  19. Ullmann, R., Schwendel, A., Klemen, H., Wolf, G., Petersen, I., and Popper, H. H. (1998) Unbalanced chromosomal aberrations in neuroendocrine lung tumors as detected by comparative genomic hybridization. Hum. Pathol. 29, 1145ā€“1159.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  20. Petersen, S., Aninat-Meyer, M., SchlĆ¼ns, K., Gellert, K., Dietel, M., and Petersen, I. (2000) Chromosomal alterations in the clonal evolution to the metastatic stage of squamous cell carcinomas of the lung. Br. J. Cancer 82, 65ā€“73.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  21. Petersen, I., Hidalgo, H., Petersen, S., SchlĆ¼ns, K., Schewe, C., Pacyna-Gengelbach, M., et al. (2000) Chromosomal imbalances in brain metastases of solid tumors. Brain Pathology 10, 395ā€“401.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  22. Goeze, A., SchlĆ¼ns, K., Wolf, G., ThƤ;sler, Z., Petersen, S., and Petersen, I. (2002) Chromosomal imbalances of primary and metastatic lung adenocarcinomas. J. Pathol. 196, 8ā€“16.

    ArticleĀ  PubMedĀ  Google ScholarĀ 

  23. Ried, T., Petersen, I., Holtgreve-Grez, H., Speicher, M. R., Schrƶck, E., du Manoir, S., and Cremer, T. (1994) Mapping of multiple DNA gains and losses in primary small cell lung carcinomas by comparative genomic hybridization. Cancer Res. 54, 1801ā€“1806.

    PubMedĀ  CASĀ  Google ScholarĀ 

  24. Moore, D. H., 2nd, Pallavicini, M., Cher, M. L., and Gray, J. W. (1997) A t-statistic for objective interpretation of comparative genomic hybridization (CGH) profiles. Cytometry 28, 183ā€“190.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  25. Tirkkonen, M., Karhu, R., Kallioniemi, O., and Isola, J. (1996) Evaluation of camera requirements for comparative genomic hybridization. Cytometry 25, 394ā€“398.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  26. du Manoir, S., Kallioniemi, O.-P., Lichter, P., Piper, J., Benedetti, P. A., Carothers, A. D., et al. (1995) Hardware and software requirements for quantitative analysis of comparative genomic hybridization. Cytometry 19, 4ā€“9.

    ArticleĀ  PubMedĀ  Google ScholarĀ 

  27. Lundsteen, C., Maahr, J., Christensen, B., Bryndorf, T., Bentz, M., Lichter, P., and Gerdes, T. (1995) Image analysis in comparative genomic hybridization. Cytometry 19, 42ā€“50.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  28. Piper, J., Rutowitz, D., Sudar, D., Kallioniemi, A., Kallioniemi, O. P., Waldman, F. M., et al. (1995) Computer image analysis of comparative genomic hybridization. Cytometry 19, 10ā€“26.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  29. Knƶsel, T., Petersen, S., Schwabe, H., SchlĆ¼ns, K., Stein, U., Schlag, P. M., et al. (2002) Incidence of chromosomal imbalances in advanced colorectal carcinomas and their metastases. Virchows Arch. 440, 187ā€“194.

    ArticleĀ  PubMedĀ  Google ScholarĀ 

  30. Alers, J. C., Rochat, J., Krijtenburg, P. J., van Dekken, H., Raap, A. K., and Rosenberg, C. (1999) Universal linkage system: an improved method for labeling archival DNA for comparative genomic hybridization. Genes Chromosomes Cancer 25, 301ā€“305.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  31. Karhu, R., Rummukainen, J., Lorch, T., and Isola, J. (1999) Four-color CGH: a new method for quality control of comparative genomic hybridization. Genes Chromosomes Cancer 24, 112ā€“118.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  32. Kirchhoff, M., Gerdes, T., Rose, H., Maahr, J., Ottesen, A. M., and Lundsteen, C. (1998) Detection of chromosomal gains and losses in comparative genomic hybridization analysis based on standard reference intervals. Cytometry 31, 163ā€“173.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

Ā© 2003 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Petersen, I. (2003). Comparative Genomic Hybridization of Human Lung Cancer. In: Driscoll, B. (eds) Lung Cancer. Methods in Molecular Medicineā„¢, vol 75. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-324-0:209

Download citation

  • DOI: https://doi.org/10.1385/1-59259-324-0:209

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-0-89603-920-9

  • Online ISBN: 978-1-59259-324-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics