Abstract
As a transcriptional regulator, the androgen receptor (AR) regulates the expression of many genes that are essential for male sexual differentiation, including the development of both normal prostate and prostate cancer. The AR acts by binding to regulatory DNA sequences found on the promoters of regulated genes. The study of AR activity on such responsive promoters is greatly facilitated by the use of the reporter gene assay, which provides a quantitative and reproducible method for studying the activity of such promoters. Among the several reporter genes that can be used, the genes encoding luciferase (Luc) and chloramphenicol acetyltransferase (CAT) have been used most widely and successfully by researchers interested in AR-regulated promoters. Such studies have led to the identification and characterization of DNA regulatory elements mediating AR activity on responsive promoters and to an improved understanding of how AR regulates the transcription process. Described in this chapter is a method by which to generate and utilize Luc and CAT reporter gene plasmids driven by the promoter of a novel androgen-regulated gene, ETV1.
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References
Wilson, J.D., Griffin, J.E., George, F.W., and Leshin, M. (1981) The role of gonadal steroids in sexual differentiation. Rec. Prog. Horm. Res. 37,1–39.
Lee, C. (1996) Role of androgen in prostate growth and regression: stromal-epithelial interaction. Prostate Suppl. 6, 52–56.
Taplin, M.E. (2007) Drug insight: role of the androgen receptor in the development and progression of prostate cancer. Nat. Clin. Pract. Oncol. 4, 236–244.
Gregory, C.W., Hamil, K.G., Kim, D., Hall. S.H., Pretlow, T.G., Mohler, J.L., et al. (1998) Androgen receptor expression in androgen-independent prostate cancer is associated with increased expression of androgen-regulated genes. Cancer Res. 58, 5718–5724.
Kato, S., Matsumoto, T., Kawano, H., Sato, T, and Takeyama, K. (2004) Function of androgen receptor in gene regulations. J. Steroid Biochem. Mol. Biol. 89–90, 627–633.
Arnone, M.I., Dmochowski, I.J., and Gache, C. (2004) Using reporter genes to study cis-regulatory elements. Methods Cell Biol. 74, 621–652.
Cai, C., Hsieh, C.L., Omwancha, J., Zheng, Z., Chen, S.Y., Baert, J.L., et al. (2007) ETV1 is a novel androgen receptor-regulated gene that mediates prostate cancer cell invasion. Mol. Endocrinol. 21, 1835–1846.
Horoszewicz, J.S., Leong, S.S., Kawinski, E., Karr, J.P., Rosenthal, H., Chu, et al. (1983) LNCaP model of human prostatic carcinoma. Cancer Res. 43, 1809–1818.
Heisler, L.E., Evangelou, A., Lew, A.M., Trachtenberg, J., Elsholtz, H.P., and Brown, T.J. (1997) Androgen-dependent cell cycle arrest and apoptotic death in PC-3 prostatic cell cultures expressing a full-length human androgen receptor. Mol. Cell. Endocrinol. 126, 59–73.
Jenster, G. (1999) The role of the androgen receptor in the development and progression of prostate cancer. Semin Oncol. 26407–421.
Jain, A., Lam, A., Vivanco, I., Carey, M.F., and Reiter, R.E. (2002) Identification of an androgen-dependent enhancer within the prostate stem cell antigen gene. Mol. Endocrinol. 16, 2323–2337.
Marques, R.B., van Weerden, W.M., Erkens-Schulze, S, de Ridder, C.M., Bangma, C.H., Trapman, J., et al. (2006) The human PC346 xenograft and cell line panel: a model system for prostate cancer progression. Eur. Urol. 49, 245–257.
Shemshedini, L., Knauthe, R., Sassone-Corsi, P., Pornon, A., and Gronemeyer, H. (1991) Cell-specific inhibitory and stimulatory effects of Fos and Jun on transcription activation by nuclear receptors. EMBO J. 10, 3839–3849.
Acknowledgments
The author would like to thank Chen-Lin Hsieh for critical reading of the manuscript. This work was supported by R15 DK067059.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Shemshedini, L. (2009). Use of Reporter Genes to Study Promoters of the Androgen Receptor. In: Park-Sarge, OK., Curry, T. (eds) Molecular Endocrinology. Methods in Molecular Biology, vol 590. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-378-7_12
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DOI: https://doi.org/10.1007/978-1-60327-378-7_12
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