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 (3–5).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Liang, P. and Pardee, A. B. (1992) Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257, 967–971.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Wang, X. and Feuerstein, G. Z. (1995) Direct sequencing of DNA isolated from mRNA differential display. Biotechniques 18, 448–453.
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.
Zhang, H., Zhang, R., and Liang, P. (1997) Differential screening of differential display cDNA products by reverse northern. Meth. Mol. Biol. 85, 87–93.
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.
Martin, K. J. and Pardee, A. B. (1999) Principles of differential display. Meth. Enzymol. 303, 234–258.
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.
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.
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.
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.
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.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights 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