Abstract
Cell-based screening methods for nuclear receptor ligands that use transgenic plant cells expressing a single human nuclear receptor (NR) may have advantages over other eukaryotic systems which express multiple NRs. For example, signal-to-noise ratio might be improved because ligands would be less likely to bind to other NRs and/or less likely to cause confounding functional changes in plant cells. As a first step toward this aim, we have expressed in plants truncated human estrogen receptor (ER) constructs linked to reporters or selective markers such as luciferase, green fluorescent protein (GFP), and hygromycin. A variety of ligands for the ER (including estradiol and known phytoestrogens) have then been tested for their ability to overexpress the linked marker gene(s) which could be measured (luciferase activity), visualized under fluorescent microscopy (GFP activity), or selected on antibiotic-containing media (hygromycin B). Our results show a close association between the effects of ER ligands in the transgenic plant roots and their effects on native ERs in mammalian cells. With the stable expression of an ERalpha-GFP ligand detection system in Arabidopsis thaliana, the estradiol-mediated response in transgenic roots is inhibited by an ER partial agonist (tamoxifen) and an antagonist (fulvestrant) at concentrations relevant to their use in breast cancer. We conclude that it is possible to express human NRs in plants in a form that can report on exogenous or endogenous ER ligands and that these constructs have a pharmacology which is relevant to ligands for the native NRs in human cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ahn KS, Sethi G, Sung B, Goel A, Ralhan R, Aggarwal BB (2008) Guggulsterone, a farnesoid X receptor antagonist, inhibits constitutive and inducible STAT3 activation through induction of a protein tyrosine phosphatase SHP-1. Cancer Res 68:4406–4415
Banerjee S, Li Y, Wang Z, Sarkar FH (2008) Multi-targeted therapy of cancer by genistein. Cancer Lett 269:226–242
Brunelli E, Minassi A, Appendino G, Moro L (2007) 8-Prenylnaringenin, inhibits estrogen receptor-alpha mediated cell growth and induces apoptosis in MCF-7 breast cancer cells. J Steroid Biochem Mol Biol 107:140–148
Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743
Connell BJ, Saleh TM (2011) Differential neuroprotection of selective estrogen receptor agonists against autonomic dysfunction and ischemic cell death in permanent versus reperfusion injury. Adv Pharm Sci 2011:1–9
De Wilde A, Lieberherr M, Colin C, Pointillart A (2004) A low dose of daidzein acts as an ERbeta-selective agonist in trabecular osteoblasts of young female piglets. J Cell Physiol 2:253–262
Demain AL (2002) Prescription for an ailing pharmaceutical industry. Nat Biotechnol 20:331
Gyling M, Leclercq G (1988) Estrogen and antiestrogen interaction with estrogen receptor of MCF-7 cells—relationship between processing and estrogenicity. J Steroid Biochem 29(1):1–8
Ikeda K, Arao Y, Otsuka H, Nomoto S, Horiguchi H, Kato S, Kayama F (2002) Terpenoids found in the umbelliferae family act as agonists/antagonists for ER(alpha) and ERbeta: differential transcription activity between ferutinine-liganded ER(alpha) and ERbeta. Biochem Biophys Res Commun 291:354–360
Levenson AS, Gehm BD, Pearce ST, Horiguchi J, Simons LA, Ward JE, Jameson JL, Jordan VC (2003) Resveratrol acts as an estrogen receptor (ER) agonist in breast cancer cells stably transfected with ER alpha. Int J Cancer 104:587–596
Louvion JF, Havaux-Copf B, Picard D (1993) Fusion of GAL4-VP16 to a steroid-binding domain provides a tool for gratuitous induction of galactose-responsive genes in yeast. Gene 131:129–134
Maggiolini M, Recchia AG, Bonofiglio D, Catalano S, Vivacqua A, Carpino A, Rago V, Rossi R, Ando S (2005) The red wine phenolics piceatannol and myricetin act as agonists for estrogen receptor alpha in human breast cancer cells. J Mol Endocrinol 35:269–281
Mersereau JE, Levy N, Staub RE, Baggett S, Zogovic T, Chow S, Ricke WA, Tagliaferri M, Cohen I, Bjeldanes LF, Leitman DC (2008) Liquiritigenin is a plant-derived highly selective estrogen receptor beta agonist. Mol Cell Endocrinol 283:49–57
Miksicek RJ (1995) Estrogenic flavonoids: structural requirements for biological activity. Proc Soc Exp Biol Med 208:44–50
Moore LB, Goodwin B, Jones SA, Wisely GB, Serabjit-Singh CJ, Willson TM, Collins JL, Kliewer SA (2000) St. John’s wort induces hepatic drug metabolism through activation of the pregnane X receptor. Proc Natl Acad Sci U S A 97:7500–7502
Moosmann B, Behl C (1999) The antioxidant neuroprotective effects of estrogens and phenolic compounds are independent from their estrogenic properties. Proc Natl Acad Sci U S A 96:8867–8872
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Pearce ST, Jordan VC (2004) The biological role of estrogen receptors alpha and beta in cancer. Crit Rev Oncol Hematol 50:3–22
Raskin I, Ribnicky DM, Komarnytsky S, Ilic N, Poulev A, Borisjuk N, Brinker A, Moreno DA, Ripoll C, Yakoby N, O’Neal JM, Cornwell T, Pastor I, Fridlender B (2002) Plants and human health in the twenty-first century. Trends Biotechnol 20:522–531
Siemering KR, Golbik R, Sever R, Haseloff J (1996) Mutations that suppress the thermosensitivity of green fluorescent protein. Curr Biol 6:1653–1663
Tavva VS, Dinkins RD, Palli SR, Collins GB (2006) Development of a methoxyfenozide-responsive gene switch for applications in plants. Plant J 45:457–469
Usui T (2006) Pharmaceutical prospects of phytoestrogens. Endocr J 53:7–20
Xu H, Kraus WL, Shuler ML (2008) Development of a stable dual cell-line GFP expression system to study estrogenic endocrine disruptors. Biotechnol Bioeng 101:1276–1287
Acknowledgments
This work was supported by the National Center for Complementary & Alternative Medicine (NIH grant R42 AT006639 awarded to John Littleton, Naprogenix Inc.).
Conflict of Interest
Authors declare that they have no conflict of interest. John Littleton and Dennis Trent Rogers are employees of Naprogenix Inc. that is the sole licensee of the technology from the University of Kentucky Research Foundation.
Ethical Statement
No human or animal samples were used during this research experiment.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Gunjan, S.K., Rogers, D.T., Zhang, J. et al. Use of Alpha-, Beta-Estrogen Receptor as a “New Tool” for Detection of Specific Small Molecule Activity. Plant Mol Biol Rep 33, 1837–1843 (2015). https://doi.org/10.1007/s11105-015-0879-5
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11105-015-0879-5