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Methods for Identifying and Studying Genetic Alterations in Hormone-Dependent Cancers

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The Nuclear Receptor Superfamily

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 505))

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Abstract

Genetic alterations underlying the development of cancer include large chromosomal aberrations, such as amplifications, deletions and translocations as well as small changes in sequence, i.e. mutations. Thus, different methods are needed to reveal various types of genetic changes. Fluorescence in situ hybridisation (FISH) is a versatile technique for detecting chromosomal alterations either in cultured cells or even in formalin-fixed paraffin-embedded tissue. For screening mutations, denaturing high-performance liquid chromatography (DHPLC) provides a relatively fast, cheap and sensitive option. The only special requirement is the HPLC equipment suitable for the analysis. As a screening tool, it does not reveal the actual base pair change, which in the end needs to be done by sequencing. FISH and DHPLC can both be utilized in research as well as in clinical diagnostic laboratories.

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References

  1. Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL. (1987) Human breast cancer: correlation of relapse and survival with amplification of the HER-2/ neu oncogene. Science 235:177–182.

    Article  CAS  PubMed  Google Scholar 

  2. Visakorpi T, Hyytinen E, Koivisto P, et al. (1995) In vivo amplification of the androgen receptor gene and progression of human prostate cancer. Nat Genet9:401–406.

    Article  CAS  PubMed  Google Scholar 

  3. Tomlins SA, Rhodes DR, Perner S, et al. (2005) Recurrent fusion of TMPRSS2 and ETS transcription factor genes in prostate cancer. Science 310:644–648.

    Article  CAS  PubMed  Google Scholar 

  4. Hollstein M, Sidransky D, Vogelstein B, Harris CC. (1991) p53 mutations in human cancers. Science 253:49–53.

    Article  CAS  PubMed  Google Scholar 

  5. Taplin ME, Bubley GJ, Shuster TD, et al. (1995) Mutation of the androgen-receptor gene in metastatic androgen-independent prostate cancer. N Engl J Med 332:1393–1398.

    Article  CAS  PubMed  Google Scholar 

  6. Hyytinen E, Visakorpi T, Kallioniemi A, Kallioniemi OP, Isola J. (1994) Improved technique for analysis of formalin fixed paraffin embedded tumors by fluorescence in situ hybridization. Cytometry 16:93–99.

    Article  CAS  PubMed  Google Scholar 

  7. Saramäki O, Willi N, Bratt O, et al. (2001) Amplification of EIF3S3 gene is associated with advanced stage in prostate cancer. Am J Pathol 159:2089–2094.

    PubMed  Google Scholar 

  8. Thomas RK, Nickerson E, Simons JF, et al. (2006) Sensitive mutation detection in heterogeneous cancer specimens by massively parallel picoliter reactor sequencing. Nat Med 12:852–855.

    Article  CAS  PubMed  Google Scholar 

  9. Ainsworth PJ, Rodenhiser DI. (1994) A nonradioactive method for the detection of single-strand conformational polymorphisms (SSCP). Methods Mol Biol 31:205–210.

    CAS  PubMed  Google Scholar 

  10. Ren J. (2000) High-throughput single-strand conformation polymorphism analysis by capillary electrophoresis. J Chromatogr B Biomed Sci Appl 741(2):115–128.

    Article  CAS  PubMed  Google Scholar 

  11. Fodde R, Losekoot M. (1994) Mutation detection by denaturing gradient gel electrophoresis (DGGE). Hum Mutat 3(2):83–94.

    Article  CAS  PubMed  Google Scholar 

  12. Xiao W, Oefner PJ. (2001) Denaturing high-performance liquid chromatography: A review. Hum Mutat 17:439–474.

    Article  CAS  PubMed  Google Scholar 

  13. Waltering KK, Wallén MJ, Tammela TL, Vessella RL, Visakorpi T. (2006) Mutation screening of the androgen receptor promoter and untranslated regions in prostate cancer. Prostate 66(15):1585–1591.

    Article  CAS  PubMed  Google Scholar 

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Saramäki, O.R., Waltering, K.K., Visakorpi, T. (2009). Methods for Identifying and Studying Genetic Alterations in Hormone-Dependent Cancers. In: McEwan, I.J. (eds) The Nuclear Receptor Superfamily. Methods in Molecular Biology™, vol 505. Humana Press. https://doi.org/10.1007/978-1-60327-575-0_15

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  • DOI: https://doi.org/10.1007/978-1-60327-575-0_15

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60327-574-3

  • Online ISBN: 978-1-60327-575-0

  • eBook Packages: Springer Protocols

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