ABSTRACT
4-Hydroxy tamoxifen (OHT) and trilostane interact differently with the oestrogen receptor (ER). OHT is a competitive inhibitor whereas trilostane has direct, but non-competitive effects on ER. This study compared the effects of OHT and trilostane, in the presence of 17beta-oestradiol (E2) on gene expression in MCF-7 breast cancer cells using microarrays each representing nearly 20,000 human genes. Striking differences between the sets of genes affected by these two drugs were observed. Both OHT and trilostane affected transcription of genes involved in cell cycle regulation, cell adhesion and matrix formation, however, only 12.5% of trilostane down-regulated genes and 9.2% of up-regulated genes were similarly regulated by OHT. A selective up-regulation of ERbeta by trilostane, but not OHT, was observed and confirmed by qRT-PCR. Similar up-regulation of this gene by trilostane was observed in the uterus of trilostane-treated (4 mg/kg for 7 days) rats, in which ERbeta mRNA (3-fold) and ERbeta protein expression (10-fold) were both increased. These data show that OHT and trilostane regulate the expression of different sets of genes, reflecting their different modes of interaction with ER. Trilostane-specific up-regulation of ERbeta could explain its positive benefit rates in acquired tamoxifen resistance.
Subject(s)
Antineoplastic Agents/pharmacology , Dihydrotestosterone/analogs & derivatives , Estrogen Antagonists/pharmacology , Estrogen Receptor beta/metabolism , Estrogens/metabolism , Gene Expression Regulation/drug effects , Tamoxifen/analogs & derivatives , Animals , Antineoplastic Agents/metabolism , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Cell Line, Tumor , Dihydrotestosterone/metabolism , Dihydrotestosterone/pharmacology , Estrogen Antagonists/metabolism , Estrogen Receptor beta/genetics , Female , Gene Expression Profiling , Humans , Molecular Sequence Data , Oligonucleotide Array Sequence Analysis , Rats , Rats, Wistar , Tamoxifen/metabolism , Tamoxifen/pharmacology , Up-RegulationABSTRACT
Currently available antioestrogens, such as tamoxifen, are competitive inhibitors that bind to the ligand binding sites of oestrogen receptors, ERalpha and ERbeta. The search for alternative anti-hormone therapies is prompted by the need for drugs that are effective when tumours become tamoxifen resistant. The existence of different receptor isoforms also raise the possibility of improving selectivity. Earlier use of the 3beta-hydroxysteroid dehydrogenase inhibitor, trilostane (4alpha,5- epoxy-17beta-hydroxy-3-oxo-5alpha-androstane-2alpha-carbonitrile), suggested that it had beneficial actions in breast cancer that were only partially attributable to inhibition of steroidogenesis. The present studies on the interactions of trilostane with oestrogen receptors show that it (i) inhibits oestrogen-stimulated proliferation in MCF-7 breast cancer cells, (ii) enhances the affinity of oestradiol binding to ER in rat uteri and specifically increases oestradiol binding to an ERbeta-like isoform, (iii) inhibits ERalpha and ERbeta binding to the classical vitellogenin gene oestrogen response element (ERE) and (iv) inhibits oestrogen-stimulated gene transcription in ERE-linked reporter systems in MCF-7 cells. The results demonstrate a novel, presumably allosteric, mode of antioestrogen action. The beneficial actions of trilostane in breast cancer may be attributed to the combination of this antioestrogen effect with its well documented suppression of steroidogenesis.
