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cDNA Microarray and Bioinformatic Analysis of Nuclear Factor-κB Related Genes in Squamous Cell Carcinoma

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Cancer Genomics and Proteomics

Part of the book series: Methods in Molecularbiology™ ((MIMB,volume 383))

Abstract

Squamous cell carcinomas and several other cancers have been found to exhibit microarray expression profiles that include genes related to nuclear factor (NF)-κB, a signal activated transcription factor that is evolutionarily important in regulating early response gene programs to injury and infection. Inhibition of NF-κB by expression of a dominant negative signal phosphorylation site mutant of inhibitor-κB, IκBαM, under a tetracycline inducible promoter, established the role of NF-κB as an essential molecular switch modulating multiple genes important in the malignant phenotype. Bioinfomatic analysis of the promoter and coding region of IκBαM-modulated genes has enabled identification of new candidates with and without known NF-κB related motifs for validation and functional studies of their relationship to NF-κB. These studies illustrate how microarray data can be used to generate a hypothesis regarding regulation of genes by a specific signal transcription factor, and how genetic mutants and bioinformatic analysis can be used to analyze the relative importance of the regulatory molecule to expression of genes involved in the malignant phenotype.

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References

  1. Dong, G., Loukinova, E., Chen, Z., et al. (2001) Molecular profiling of transformed and metastatic murine squamous carcinoma cells by differential display and cDNA microarray reveals altered expression of multiple genes related to growth, apoptosis, angiogenesis, and the NF-kappaB signal pathway. Cancer Res. 61, 4797–4808.

    CAS  PubMed  Google Scholar 

  2. Davis, R. E., Brown K. D., Siebenlist, U., and Staudt, L. M. (2001) Constitutive nuclear factor kappaB activity is required for survival of activated B cell-like diffuse large B cell lymphoma cells. J. Exp. Med. 194, 1861–1874.

    Article  CAS  PubMed  Google Scholar 

  3. Hinz. M., Lemke, P., Anagnostopoulos, I., et al. (2002) Nuclear factor kappaB-dependent gene expression profiling of Hodgkin’s disease tumor cells, pathogenetic significance, and link to constitutive signal transducer and activator of transcription 5a activity. J. Exp. Med. 196, 605–617.

    Article  CAS  PubMed  Google Scholar 

  4. Loercher, A., Lee, T. L., Ricker, J. L., et al. (2004) Nuclear factor-kappaB is an important modulator of the altered gene expression profile and malignant phenotype in squamous cell carcinoma. Cancer Res. 64, 6511–6523.

    Article  CAS  PubMed  Google Scholar 

  5. Chen, Z., Smith, C.W., Kiel, D., and Van Waes, C. (1997) Metastatic variants derived following in vivo tumor progression of an in vitro transformed squamous cell carcinoma line acquire a differential growth advantage requiring tumor-host interaction. Clin. Exp. Metastasis 15, 527–537.

    Article  CAS  PubMed  Google Scholar 

  6. Dong, G., Chen, Z., Kato, T., and Van Waes, C. (1999) The host environment promotes the constitutive activation of nuclear factor-kappaB and proinflammatory cytokine expression during metastatic tumor progression of murine squamous cell carcinoma. Cancer Res. 59, 3495–3504.

    CAS  PubMed  Google Scholar 

  7. Dennis, G., Jr., Sherman, B. T., Hosack, D. A., et al. (2003) DAVID: database for Annotation, Visualization, and Integrated Discovery. Genome Biol. 4, 9R60. 1–11 (http://genomebiology.com/2003/4/9/R60).

    Article  Google Scholar 

  8. Krause, C. J., Carey, T. E., Ott, R. W., Hurbis, C., McClatchey, K. D., and Regezi J. A. (1981) Human squamous cell carcinoma. Establishment and characterization of new permanent cell lines. Arch. Otolaryngol. 107, 703–710.

    CAS  PubMed  Google Scholar 

  9. Chen, Y., Dougherty, E. R., and Bittner, M. L. (1997) Ratio-based decisions and the quantitative analysis of cDNA microarray images. J. Biomed. Optics 2, 364–374.

    Article  CAS  Google Scholar 

  10. Chung, C. H., Parker, J. S., Karaca, G., et al. (2004) Molecular classification of head and neck squamous cell carcinomas using patterns of gene expression. Cancer Cell 5, 489–500.

    Article  CAS  PubMed  Google Scholar 

  11. Eisen, M. B., Spellman, P. T., Brown, P. O., and Botstein, D. (1998) Cluster analysis and display of genome-wide expression patterns. Proc. Natl. Acad. Sci. USA 95, 14,863–14,868.

    Article  CAS  PubMed  Google Scholar 

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© 2007 Humana Press Inc., Totowa, NJ

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Chen, Z., Lee, TL., Yang, XP., Dong, G., Loercher, A., Van Waes, C. (2007). cDNA Microarray and Bioinformatic Analysis of Nuclear Factor-κB Related Genes in Squamous Cell Carcinoma. In: Fisher, P.B. (eds) Cancer Genomics and Proteomics. Methods in Molecularbiology™, vol 383. Humana Press. https://doi.org/10.1007/978-1-59745-335-6_6

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  • DOI: https://doi.org/10.1007/978-1-59745-335-6_6

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-504-0

  • Online ISBN: 978-1-59745-335-6

  • eBook Packages: Springer Protocols

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