Skip to main content

Detection of Differentially Expressed Genes in Cancer Using Differential Display

  • Protocol
Molecular Analysis of Cancer

Part of the book series: Methods in Molecular Medicine ((MIMM,volume 68))

  • 340 Accesses

Abstrect

It is estimated that the human genome contains 25,000–35,000 genes; however, only about 15% of these are expressed in any given cell and different cells and tissues express different gene subsets. Gene expression is tightly regulated under normal physiologic conditions but is often disrupted during malignant transformation and tumor progression. Analysis of differential gene expression in such events as cancer is essential to better understand these complicated processes so that new diagnoses and forms of treatment can be developed. RNA differential display technology offers a straightforward and efficient way to detect differential gene expression in a variety of physiologic and pathologic circumstances including cancer (1,2). By comparing messenger RNA expression patterns between tumor cells and their normal counterparts on the same gel, altered gene expression in tumor cells can be easily detected. This technique has been widely and successfully applied not only in cancer research but also in many other areas of biologic research and has resulted in thousands of publications in the few years since it was developed (35).

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. Liang, P. and Pardee, A. B. (1992) Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257, 967–971.

    Article  PubMed  CAS  Google Scholar 

  2. Liang, P., Bauer, D., Averboukh, L., Warthoe, P., Rohrwild, M., Muller, H., et al. (1995) Analysis of altered gene expression by differential display. Meth. Enzymol. 254, 304–321.

    Article  Google Scholar 

  3. Fu, Y., Comella, N., Tognazzi, K., Brown, L. F., Dvorak, H. F., and Kocher, O. (1999) Cloning of DLM-1, a novel gene that is up-regulated in activated macrophages, using RNA differential display. Gene 240, 157–163.

    Article  PubMed  CAS  Google Scholar 

  4. Guan, R. J., Ford, H. L., Fu, Y., Li, Y., Shaw, L. M., and Pardee, A. B. (2000) Drg-1 as a differentiation-related, putative metastatic suppressor gene in human colon cancer. Cancer Res. 60, 749–755.

    PubMed  CAS  Google Scholar 

  5. Silva, I. D., Salicioni, A. M., Russo, I. H., Higgy, N. A., Gebrim, L. H., and Russo, J. (1997) Tamoxifen down-regulates CD36 messenger RNA levels in normal and neoplastic human breast tissues. Cancer Res. 57, 378–381.

    PubMed  CAS  Google Scholar 

  6. St. John, T. P. and Davis, R. W. (1979) Isolation of galactose-inducible DNA sequences from Saccharomyces cerevisiae by differential plaque filter hybridization. Cell 16, 443–452.

    Article  Google Scholar 

  7. Zimmermann, C. R., Orr, W. C., Leclerc, R. F., Barnard, E. C., and Timberlake, W. E. (1980) Molecular cloning and selection of genes regulated in Aspergillus development. Cell 21, 709–715.

    Article  PubMed  CAS  Google Scholar 

  8. 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, 467–470.

    Article  PubMed  CAS  Google Scholar 

  9. Liang, P., Averboukh, L., and Pardee, A. B. (1993) Distribution and cloning of eukaryotic mRNAs by means of differential display: refinements and optimization. Nucleic Acids Res. 21, 3269–3275.

    Article  PubMed  CAS  Google Scholar 

  10. Li, F., Barnathan, E. S., and Kariko, K. (1994) Rapid method for screening and cloning cDNAs generated in differential mRNA display: application of northern blot for affinity capturing of cDNAs. Nucleic Acids Res. 22, 1764, 1765.

    Article  PubMed  CAS  Google Scholar 

  11. Martin, K. J., Kwan, C. P., O’Hare, M. J., Pardee, A. B., and Sager, R. (1998) Identification and verification of differential display cDNAs using gene-specific primers and hybridization arrays. Biotechniques 6, 1018–1026.

    Google Scholar 

  12. Kocher, O., Cheresh, P., Brown, L. F., and Lee, S. W. (1995) Identification of a novel gene, selectively up-regulated in human carcinomas, using the differential display technique. Clin. Cancer Res. 1, 1209–1215.

    PubMed  CAS  Google Scholar 

  13. Chuaqui, R. F., Englert, C. R., Strup, S. E., Vocke, C. D., Zhuang, Z., Duray, P. H., et al. (1997) Identification of a novel transcript up-regulated in a clinically aggressive prostate carcinoma. Urology 50, 302–307.

    Article  PubMed  CAS  Google Scholar 

  14. Chakravarty, G., Roy, D., Gonzales, M., Gay, J., Contreras, A., and Rosen, J. M. (2000) P190-B, a Rho-GTPase-activating protein, is differentially expressed in terminal end buds and breast cancer. Cell Growth Differ. 11, 343–354.

    PubMed  CAS  Google Scholar 

  15. Schutze, K., Posl, H., and Lahr, G. (1998) Laser micromanipulation systems as universal tools in cellular and molecular biology and in medicine. Cell Mol. Biol. 44, 735–746.

    PubMed  CAS  Google Scholar 

  16. Suarez-Quian, C. A., Goldstein, S. R., Pohida, T., Smith, P. D., Peterson, J. I., Wellner, E., et al. (1999) Laser capture microdissection of single cells from complex tissues. Biotechniques 26, 328–335.

    PubMed  CAS  Google Scholar 

  17. Wang, X. and Feuerstein, G. Z. (1995) Direct sequencing of DNA isolated from mRNA differential display. Biotechniques 18, 448–453.

    PubMed  CAS  Google Scholar 

  18. Mou, L., Miller, H., Li, J., Wang, E., and Chalifour, L. (1994) Improvements to the differential display method for gene analysis. Biochem. Biophys. Res. Commun. 199, 564–569.

    Article  PubMed  CAS  Google Scholar 

  19. Zhang, H., Zhang, R., and Liang, P. (1997) Differential screening of differential display cDNA products by reverse northern. Meth. Mol. Biol. 85, 87–93.

    CAS  Google Scholar 

  20. Callard, D., Lescure, B., and Mazzolini, L. (1994) A method for the elimination of false positives generated by the mRNA differential display technique. Biotechniques 16, 1096–1103.

    PubMed  CAS  Google Scholar 

  21. Martin, K. J. and Pardee, A. B. (1999) Principles of differential display. Meth. Enzymol. 303, 234–258.

    Article  PubMed  CAS  Google Scholar 

  22. Kalman, M., Kalman, E. T., and Cashel, M. (1990) Polymerase chain reaction (PCR) amplification with a single specific primer. Biochem. Biophys. Res. Commun. 167, 504–506.

    Article  PubMed  CAS  Google Scholar 

  23. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Extraction, purification, and analysis of messenger RNA from eukaryotic cells, in Molecular Cloning, A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Plainview, NY, pp. 7.2–7.87.

    Google Scholar 

  24. Donohue, P. J., Alberts, G. F., Guo, Y., and Winkles, J. A. (1995) Identification by targeted differential display of an immediate early gene encoding a putative serine/threonine kinase. J. Biol. Chem. 27010, 351–10,357.

    Google Scholar 

  25. Haag, E. and Raman, V. (1994) Effects of primer choice and source of Taq DNA polymerase on the banding patterns of differential display RT-PCR. BioTechniques 17, 226–228.

    PubMed  CAS  Google Scholar 

  26. Zimmermann, J. W. and Schultz, R. M. (1994) Analysis of gene expression in the preimplantation mouse embryo: use of mRNA differential display. Proc. Natl. Acad. Sci. USA 91, 5456–5460.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2002 Humana Press Inc.

About this protocol

Cite this protocol

Fu, Y. (2002). Detection of Differentially Expressed Genes in Cancer Using Differential Display. In: Boultwood, J., Fidler, C. (eds) Molecular Analysis of Cancer. Methods in Molecular Medicine, vol 68. Humana Press. https://doi.org/10.1385/1-59259-135-3:179

Download citation

  • DOI: https://doi.org/10.1385/1-59259-135-3:179

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-622-2

  • Online ISBN: 978-1-59259-135-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics