Printer Friendly

Mechanisms of endocrine resistance.


Two-thirds of breast cancers are associated with oestrogen receptor (ER) signalling, and hormonal manipulation was first shown to be a successful strategy against breast cancer over 100 years ago [1]. Currently available endocrine strategies include targeting the ER itself with the selective ER modulator tamoxifen or the ER downregulator fulvestrant, as well as suppressing the amount of available ligand for the receptor with gonadal suppression in premenopausal women or aromatase inhibitors (AIs) in postmenopausal women.

Among postmenopausal women with early ER-positive breast cancer, endocrine therapy has been shown to have a greater impact on reducing annual breast cancer death rate than adjuvant chemotherapy (31% versus 20%) [2]. Given their favourable side effect profile and their potential for significant anti-tumour activity, endocrine therapies represent an important tool in our armamentarium against breast cancer. Unfortunately, despite documented levels of ERs, up to 50% of patients with metastatic disease do not respond to first-line endocrine treatment (de novo resistance), while the remainder will eventually relapse (acquired resistance) [3]. Understanding the various biological mechanisms responsible for the development of endocrine resistance can identify new strategies to enhance the efficacy of hormone receptor-positive breast cancer treatment.

A number of theories, supported by preclinical data, have been proposed to explain endocrine resistance. These vary according to the sustained dependence on ER-mediated signalling and a significant number implicate growth factor-mediated mitogenic signalling (Table 1). Identifying which resistance mechanism(s) is (are) operational is of obvious clinical relevance in tailoring the most appropriate subsequent therapy (i.e. non-ER-targeted treatment, further endocrine manipulation, or a combination of both).

Normal ER function

The ER belongs to the steroid nuclear receptor family of ligand-dependent transcription factors (Figure 1A). The transcription of ER-regulated genes is regulated by two activating function (AF) domains. AF-2 is located at the C-terminus and requires binding of oestradiol (E2) to the ligand-binding domain (LBD) of the receptor, whereas AF-1 is located at the N-terminus and is hormone independent. E2 binding to the LBD induces a conformational change in the receptor, leading to the recruitment of nuclear co-activators (NCOAs) that increase ER interaction with DNA [4]. The activated ER binds as a dimer to oestrogen response elements in the promoters of target genes and activates transcription. In contrast, AF-1 can be phosphorylated by growth factor receptors or other downstream effectors and can act both independently or synergistically with AF-2 to increase the efficiency of ER transcriptional activity [5].


Tamoxifen acts as a competitive inhibitor. Upon binding to the LBD, it induces a conformational change that inactivates AF-2, but has no effect on AF-1 transcriptional activity [6]. Because ER activity in the breast is predominantly mediated via AF-2, tamoxifen has an overall antagonist effect in the breast, but can act as an agonist in other tissues primarily driven by AF-1, such as the uterus [7].

In addition, tamoxifen recruits co-repressors (NCORs) thereby further inhibiting transcription. Alterations in the relative contributions of AF-1/AF-2 to transcription, as well as the amounts of NCOAs/NCORs available, may shift the balance of tamoxifen activity towards agonism and have been implicated in endocrine resistance. In addition to its role as a classical transcription factor, ER can also enhance transcription without direct DNA binding by participating in protein-protein interactions with other transcription factors (non-classical genomic activity). For example, ER can essentially act as a co-activator and increase the activity of the Jun/Fos activator protein 1 (AP-1) transcription complex [8] (Figure 1B).

Finally, in addition to these nuclear transcriptional functions, non-genomic actions have more recently been ascribed to the ER. Oestrogen-bound ER has been shown to interact directly with, and phosphorylate, membrane-associated growth factor receptors [9,10] as well as downstream effectors such as the p85 subunit of phosphoinositide-3-kinase (PI3K) [11], resulting in further pro-survival and anti-apoptotic signalling (Figure 1C).

The degree to which these individual functions of the ER contribute to malignant proliferation may vary from one patient to another and evolve over time thereby contributing to the development of endocrine resistance.

Implication of growth factor receptor and downstream mitogenic signalling in endocrine resistance

Membrane growth factor receptors such as the epidermal growth factor receptor (EGFR), the human epidermal growth factor receptor 2 (HER2) or the insulin growth factor 1 receptor (IGF1R) have been implicated in endocrine resistance. Upregulation of HER2 occurs in approximately 15-20% of breast cancers and has been associated with poor prognosis and de novo resistance to tamoxifen in the neoadjuvant setting [12]. Similarly, EGFR is overexpressed in a number of breast cancers and has also been associated with poor response to tamoxifen [12]. It is important to note that cell models of acquired resistance to both tamoxifen and oestrogen deprivation (ED) have shown that the development of resistance is associated with adaptative upregulation of growth factor signalling pathways [13,14].

Activation of these membrane receptors stimulates two major intracellular kinase signalling cascades--the ras/mitogenic-activated protein kinase (MAPK) pathway and the PI3K/Akt pathway. These pathways activate downstream effectors leading to a cascade of signals involved in malignant growth and survival, and can be involved in endocrine resistance by a number of mechanisms including downregulation of ER expression, a switch to ER-independent growth, or bi-directional crosstalk between ER and mitogenic signalling.

ER downregulation

ER and HER2 expression are frequently described as inversely related. ER-positive cell lines stably transfected with full-length HER2 demonstrate downregulation of ER, while quantitative measurements of ER levels in tumour samples show consistently lower levels of the receptor among patients with HER2-amplified breast cancer [15].

There is some evidence that hyperactive growth factor signalling can suppress ER expression, and may eventually lead to complete loss of ER [16]. Conversely, interruption of hyperactive MAPK or EGFR has been shown to re-induce ER expression in cell lines and xenograft models [17,18]. In fact, in a small study of 10 patients with ER-negative/HER2-positive breast cancer receiving trastuzumab, three patients acquired ER expression with treatment [19]. Studies with the dual EGFR/HER2 inhibitor lapatinib have shown that long-term treatment with this agent was associated with adaptive increase in ER signalling [20]. The degree to which complete loss of ERs secondary to mitogenic signalling may contribute to acquired resistance needs further confirmation as serial clinical samples from women who have relapsed on endocrine therapy suggest that this is a rare event (10-15%) [21,22]. However, this 'yin and yang' dynamic interaction between ER and growth factor signalling would support using ER- and growth factor-targeted therapies in combination, or in fact in sequence, as one may sensitise to the other.

Switch to ER-independent growth

Other mechanisms are likely to account for endocrine resistance in HER2-amplified breast cancer, as measurable levels of ERs are expressed in 50% of cases. Another possible mode of escape from the inhibitory effects of endocrine therapy is an adaptive switch to ER-independent growth, where the ER is still present but no longer driving proliferation. A cell model for acquired resistance to fulvestrant has shown that cells adapt to long-term treatment with the ER downregulator by pronounced upregulation of multiple growth-stimulatory pathways, resulting in oestrogen-independent autocrine-regulated proliferation [9]. This would imply pan-endocrine resistance and support the use of non-ER-targeted therapies.

Endocrine resistance is usually associated with a functional ER

Most in vitro and clinical observations would suggest that even following the development of endocrine resistance, ER signalling continues to play an important role in the proliferation of breast cancer. Biopsies of tumours from breast cancer patients who have relapsed while receiving treatment with an anti-oestrogen agent show a functional ER [23], while women who have become refractory to tamoxifen or non-steroidal AIs actually respond to further endocrine manipulation with the ER[alpha] downregulator fulvestrant [24,25], indicating that ER-mediated signalling remains functional. There is increasing evidence that crosstalk with growth factor and downstream mitogenic pathways can augment the genomic and non-genomic functions of ERs.

Synergistic interaction between ER and mitogenic signalling resulting in enhanced genomic and non-genomic functions of ER

Increase in classical transcription via AF-1

Growth factor-mediated activation of MAPK or Akt can potentiate E2-mediated ER classical transcriptional activity by directly phosphorylating AF-1 [26]. Of note, both MAPK and Akt have been shown to phosphorylate ER within AF-1 (at serine 118 and serine 167, respectively) in the absence of E2, thereby contributing to ligand-independent ER transactivation [27,28]. Other downstream effectors of peptide growth factor receptors have also been shown to activate ERs, such as protein kinase A and the cyclin E-cdk-2 complex [29].

An increase in AF-1 transcription may predict for differential sensitivity to endocrine agents. Tamoxifen may efficiently block AF-2-mediated transcription; however, in tumours that exhibit high levels of AF-1 activity driven by mitogenic signalling, tamoxifen may act as an agonist [7]. This is supported by the high rate of primary tamoxifen resistance observed in neoadjuvant trials of HER2-amplified breast cancer [12]. There are some data to suggest that E2 deprivation using an AI, or ER downregulation using fulvestrant, may therefore be a more effective anti-cancer strategy in EGFR-positive/HER2-positive breast cancer [12,30,31].

Potentiation of genomic activity by altering the balance of co-regulators

ER transcription is tightly regulated by the balance of NCOAs/NCORs. Mitogenic signalling can alter the expression of some of these co-regulators thereby enhancing classical and non-classical ER transcription. The co-activator NCOA3, also known as amplified in breast cancer-1 (AIB1) is overexpressed in 50% of breast cancers and amplified in 5% [32]. Among untreated patients with ER-positive breast cancer it is associated with improved survival. In contrast, AIB1 has been associated with poor outcome on tamoxifen in HER2-amplified breast cancer because HER2 activates AIB1 and enhances the agonist effects of tamoxifen [33,34]. Similarly, decreased levels of NCORs have been shown to enhance tamoxifen agonism by shifting the balance towards ER transcriptional activity [35].

Cyclin D1 is overexpressed in 50% and amplified in 25% of breast cancers. As a transducer of both ER and growth factor-mediated cell cycle progression, cyclin D1 emerged as a potentially useful target in endocrine-resistant breast cancer. In addition, more recent data suggested that cyclin D1 could interact directly with ERs via recruitment of members of the steroid receptor co-activators (Src) family of NCOAs in the absence of endogenous ligand [36]. However, there are conflicting clinical data regarding the causal relationship between cyclin D1 and endocrine responsiveness [37,38].

In addition to binding directly to DNA and increasing transcription of ER-dependent genes (classical), ligand-bound ER can also complex with other transcriptional factors, such as Jun/Fos via AP-1 (non-classical genomic activity). Acquired tamoxifen resistance in tumour samples has been associated with increased AP-1 activity presumably via tamoxifen-liganded ER agonist activity [39]. The stress-activated kinase p38 MAPK is a major upstream modulator of AP-1 transcriptional activity and may potentiate the agonist effects of tamoxifen at AP-1 [40].

There are data to suggest that mitogenic signalling can alter the ratio of NCOAs/NCORs and result in a hypersensitive response of ERs to E2, or an agonist effect of tamoxifen-bound ERs. Whether profiling tumours by measuring the levels of various transcription co-regulators may offer useful predictive information regarding endocrine responsiveness has not been clearly established.

Enhanced non-genomic functions of ER

Conversely, as detailed in Figure 1, in addition to its effects on transcription, oestrogen-bound ER has also been shown to result in non-genomic effects via rapid activation of EGFR [9], IGF1R [10], HER2 [41] or the cleavage of membrane-bound growth factor receptor ligands such as epidermal growth factor or transforming growth factor [alpha] [13]. This bi-directional interaction between ERs and growth factor pathways creates a self-reinforcing synergistic loop that potentiates pro-survival signals and may allow breast cancer to escape normal endocrine responsiveness. Furthermore, the extranuclear functions of ERs appear to require a ligand, and both E2 and tamoxifen can act as agonists [9].

It is important to note that this crosstalk does not seem to be operational in hormone-sensitive ER-positive MCF7 cells; its relevance appears to be limited to HER2-amplified cell lines or those with acquired endocrine resistance [42]. Unfortunately, while supported by extensive in vitro models, the clinical relevance of the non-genomic actions of the ERs remains controversial as membrane and/or cytoplasmic ERs have yet to be conclusively demonstrated in clinical samples.

Clinical implications

Growth factor signalling has been extensively implicated in endocrine resistance. In some cases the interaction between ER and mitogenic pathways can be described as a dynamic inverse relationship, where inhibition of one results in a compensatory increase in the other. This is supported by pre-clinical and clinical data which show that growth factor inhibition may increase ER expression or function and re-sensitise breast cancer cells to endocrine therapy, and which would support combination, or in fact sequential treatment.

Alternatively, growth factor signalling can interact synergistically with the ER and augment both genomic and non-genomic functions of the receptor. This would provide a strong rationale for simultaneous blockade of both ERs and mitogenic pathways. Given this body of evidence, a number of trials have been completed exploring the benefit of various inhibitors of growth factor receptors or downstream effectors [e.g. mammalian target of rapamycin (mTOR)] in modulating endocrine resistance (for reviews see [43,44]). As shown in Table 2, despite strong pre-clinical evidence, some of these studies have yielded disappointing results, which may be in part attributable to a poor selection of patients. It is unlikely that patients will respond to a combination of specific inhibitors unless the intended target is a significant driver of endocrine-resistant growth. Conversely, while significant overexpression of HER2 is a known requirement for benefit from trastuzumab, further studies are needed to determine whether more moderate receptor expression or activation may be relevant in the setting of endocrine resistance. Most ongoing studies are therefore including parallel biological studies in an effort to define the subset of patients most likely to benefit from specific combinations (Table 3).

Other mechanisms implicated in endocrine resistance

Loss of ER expression due to promoter Hypermethylation

ER expression is an obligate, albeit insufficient, requirement for sensitivity to endocrine therapy, and loss of ERs with progressive disease due to clonal selection could account for acquired endocrine resistance. ER silencing as a result of promoter hypermethylation has been documented in a proportion of breast cancers [45]. It is important to note that this process has been shown to be reversible. Demethylating agents or histone deacetylase (HDAC) inhibitors can re-activate expression of a functional ER in cell lines with ER silencing due to promoter methylation [46,47]. These observations are provocative and have obvious clinical implications for a proportion of patients with ER-negative tumours who may benefit from endocrine therapy if ER expression could be reactivated using a demethylating agent. A trial of tamoxifen in combination with an HDAC inhibitor in patients who have relapsed after endocrine therapy is ongoing to investigate whether the HDAC inhibitor may restore endocrine sensitivity by enhancing ER expression.

Adaptive increase in ER and E2 hypersensitivity

The biological mechanisms contributing resistance to ED using AIs or gonadal suppression are less well defined than those underpinning tamoxifen resistance and may in fact be quite distinct. Long-term ED can be associated with adaptive aberrant mitogenic signalling as previously described. However, as ED strategies do not involve direct interaction with ERs, none of the paradoxical agonist activities of tamoxifen would be expected. In addition, whereas acquired tamoxifen resistance has been frequently associated with relative decrease in ER expression, adaptation to ED may lead to upregulation of ERs. In vitro models of long-term ED have shown that part of the adaptive process involves an increase in ER expression and E2 hypersensitivity to very low levels of residual oestrogen [14]. This is supported by a recent clinical trial showing that women who have relapsed on an AI respond to more potent endocrine manipulation using the irreversible AI exemestane, or ER downregulation with fulvestrant [25]. If part of the adaptive process to AIs involves E2 hypersensitivity, then there may be a rationale for continued ED in combination with a novel agent, rather than switching to it. The current Study of Faslodex, Exemestane and Arimidex (SOFEA) trial will address this question. In this trial, women who have relapsed on a non-steroidal AI (anastrozole or letrozole) are randomly assigned to the irreversible steroidal AI exemestane, fulvestrant or fulvestrant in combination with anastrozole.

ER mutations

A single activating mutation in the ER gene has been identified and shown to result in constitutive ER transactivation [48]. However, screening of clinical samples has revealed that mutations occur in fewer than 1% of breast cancers and most are silent [3,49]. In addition, a significant number of naturally occurring mRNA splice variants encoding truncated ER have been identified; however, these have not been shown to modulate endocrine responsiveness in the clinical setting [49].

Altered drug metabolism and cellular clearance

Measurements of intratumoral levels of tamoxifen have shown that women with acquired resistance to tamoxifen have lower levels than sensitive controls [50]. Whether low intracellular levels of tamoxifen are attributable to decreased influx of the drug or increased efflux via a membrane pump such as p-glycoprotein has not been established. However, this mechanism is likely to be a minor contributor to tamoxifen resistance as clinical samples have consistently demonstrated that tamoxifen saturates ERs with greater than 99.9% occupancy [51]. The cytochrome P450 2D6 enzyme is required to convert tamoxifen to its more potent metabolite, endoxifen. CYP2D6 variants have been identified and bi-allelic polymorphism results in decreased levels of circulating endoxifen, fewer hot flushes and an increased risk of relapse among women treated with tamoxifen [52]. Similarly, CYP2D6 inhibitors, such as the selective serotonin reuptake inhibitor antidepressants, which are frequently used to treat postmenopausal hot flushes, also decrease endoxifen levels leading some to suggest that these agents should be avoided in women treated with tamoxifen [52,53]. Whether women with homozygous CYP2D6 variants would benefit from higher doses of tamoxifen or an alternative endocrine therapy has not been investigated.

Alterations in aromatase expression or function

In postmenopausal women, the only source of E2 is from the aromatisation of adrenal androgens. While peripheral conversion in adipose tissue contributes to measurable levels of circulating E2, local production via tumoural aromatase activity results in 10- to 20-fold higher E2 concentrations in the tumour than in plasma [54]. Variations in tumour aromatase levels could therefore contribute to responsiveness to AIs. A small study suggested that the level of intratumoral aromatase activity could predict for response to the first-generation AI aminoglutethemide [55]. However, more recent studies have shown no correlation between aromatase mRNA levels and response to AIs [56]. A number of single-nucleotide polymorphisms (SNPs) have been identified in the aromatase gene (CYP19); however, most do not translate into a clinically significant variation in circulating E2 levels [57]. One SNP has been shown in vitro to reduce affinity of the aromatase enzyme for exemestane [58]; however, there is no clinical evidence to date that genetic variations in CYP19 lead to resistance to aromatase inhibition in vivo.


A number of theories involving ER mutations or drug metabolism and clearance have been proposed to contribute to endocrine responsiveness. However, there is an increasing body of evidence implicating growth factor receptor and downstream kinases in both de novo and acquired endocrine resistance. The nature of the interaction between ER and mitogenic signalling probably varies over time and from one patient to another. In some models, activated growth factor-mediated signalling suppresses expression of ERs and function, raising the intriguing possibility that growth factor-targeted therapy may, over time, restore endocrine responsiveness. In other cases, ERs and growth factor signalling may interact synergistically providing the basis for combination strategies. Importantly, a significant proportion of endocrine-resistant breast cancer still depends on a functional ER. Acquired resistance can be attributed to tamoxifen agonism due to AF-1-driven transcription, or an increase in non-genomic ER activity. Degrading the receptor with fulvestrant might provide an effective strategy in this setting, while ED would be expected to abrogate ligand-dependent non-genomic activity. Comparatively little is known about the mechanisms underpinning resistance to ED; however, as more postmenopausal women are receiving first-line adjuvant treatment with AIs, this question has become increasingly relevant. Both growth factor signalling and E2 hypersensitivity have been shown to contribute and would suggest that continued ED with an AI might be a superior approach to growth factor targeting alone. A huge number of trials (Table 3) are currently exploring the benefit of various targeted agents in combination or in sequence with endocrine therapy and include biological analyses that may shed further light on the clinically relevant mechanisms of endocrine resistance.


[1.] Beaston GT. On the treatment of inoperable cases of carcinoma of the mamma: suggestions for a new method of treatment with illustrative cases. Lancet, 1896, ii, 104-107.

[2.] Early Breast Cancer Trialists' Collaborative Group (EBCTCG). Effects of chemotherapy and hormonal therapy for early breast cancer on recurrence and 15-year survival: an overview of the randomised trials. Lancet, 2005, 365, 1687-1717.

[3.] Ring A and Dowsett M. Mechanisms of tamoxifen resistance. Endocr Relat Cancer, 2004, 11, 643-658.

[4.] Shibata H, Spencer TE, Onate SA et al. Role of co-activators and co-repressors in the mechanism of steroid/thyroid receptor action. Recent Prog Horm Res, 1997, 52, 141-164; discussion 164-165.

[5.] Glaros S, Atanaskova N, Zhao C et al. Activation function-1 domain of estrogen receptor regulates the agonistic and antagonistic actions of tamoxifen. Mol Endocrinol, 2006, 20, 996-1008.

[6.] Metzger D, Losson R, Bornert JM et al. Promoter specificity of the two transcriptional activation functions of the human oestrogen receptor in yeast. Nucleic Acids Res, 1992, 20, 2813-2817.

[7.] Tzukerman MT, Esty A, Santiso-Mere D et al. Human estrogen receptor transactivational capacity is determined by both cellular and promoter context and mediated by two functionally distinct intramolecular regions. Mol Endocrinol, 1994, 8, 21-30.

[8.] Kushner PJ, Agard DA, Greene GL et al. Estrogen receptor pathways to AP-1. J Steroid Biochem Mol Biol, 2000, 74, 311-317.

[9.] Fan P, Wang J, Santen RJ et al. Long-term treatment with tamoxifen facilitates translocation of estrogen receptor alpha out of the nucleus and enhances its interaction with EGFR in MCF-7 breast cancer cells. Cancer Res, 2007, 67, 1352-1360.

[10.] Kahlert S, Nuedling S, van Eickels M et al. Estrogen receptor alpha rapidly activates the IGF-1 receptor pathway. J Biol Chem, 2000, 275, 18447-18453.

[11.] Simoncini T, Hafezi-Moghadam A, Brazil DP et al. Interaction of oestrogen receptor with the regulatory subunit of phosphatidylinositol-3-OH kinase. Nature, 2000, 407, 538-541.

[12.] Ellis MJ, Coop A, Singh B et al. Letrozole is more effective neoadjuvant endocrine therapy than tamoxifen for ErbB-1-and/or ErbB-2-positive, estrogen receptor-positive primary breast cancer: evidence from a phase III randomized trial. J Clin Oncol, 2001, 19, 3808-3816.

[13.] Knowlden JM, Hutcheson IR, Jones HE et al. Elevated levels of epidermal growth factor receptor/c-erbB2 heterodimers mediate an autocrine growth regulatory pathway in tamoxifen-resistant MCF-7 cells. Endocrinology, 2003, 144, 1032-1044,

[14.] Martin LA, Farmer I, Johnston SR et al. Enhanced estrogen receptor (ER) alpha, ERBB2, and MAPK signal transduction pathways operate during the adaptation of MCF-7 cells to long term estrogen deprivation. J Biol Chem, 2003, 278, 30458-30468.

[15.] Konecny G, Pauletti G, Pegram M et al. Quantitative association between HER-2/neu and steroid hormone receptors in hormone receptor-positive primary breast cancer. J Natl Cancer Inst, 2003, 95, 142-153.

[16.] Creighton CJ, Hilger AM, Murthy S et al. Activation of mitogen-activated protein kinase in estrogen receptor alpha-positive breast cancer cells in vitro induces an in vivo molecular phenotype of estrogen receptor alpha-negative human breast tumors. Cancer Res, 2006, 66, 3903-3911,

[17.] Oh AS, Lorant LA, Holloway JN et al. Hyperactivation of MAPK induces loss of ERalpha expression in breast cancer cells. Mol Endocrinol, 2001, 15, 1344-1359.

[18.] Massarweh S, Osborne CK, Jiang S et al. Mechanisms of tumor regression and resistance to estrogen deprivation and fulvestrant in a model of estrogen receptor-positive, HER-2/neu-positive breast cancer. Cancer Res, 2006, 66, 8266-8273.

[19.] Munzone E, Curigliano G, Rocca A et al. Reverting estrogen-receptor-negative phenotype in HER-2-overexpressing advanced breast cancer patients exposed to trastuzumab plus chemotherapy. Breast Cancer Res, 2006, 8, R4.

[20.] Xia W, Bacus S, Hegde P et al. A model of acquired autoresistance to a potent ErbB2 tyrosine kinase inhibitor and a therapeutic strategy to prevent its onset in breast cancer. Proc Natl Acad Sci U S A, 2006, 103, 7795-7800.

[21.] Johnston SR, Saccani-Jotti G, Smith IE et al. Changes in estrogen receptor, progesterone receptor, and pS2 expression in tamoxifen-resistant human breast cancer. Cancer Res, 1995, 55, 3331-3338.

[22.] Kuukasjarvi T, Kononen J, Helin H et al. Loss of estrogen receptor in recurrent breast cancer is associated with poor response to endocrine therapy. J Clin Oncol, 1996, 14, 2584-2589.

[23.] Johnston SR, Lu B, Dowsett M et al. Comparison of estrogen receptor DNA binding in untreated and acquired antiestrogen-resistant human breast tumors. Cancer Res, 1997, 57, 3723-3727.

[24.] Dowsett M, Nicholson RI and Pietras RJ. Biological characteristics of the pure antiestrogen fulvestrant: overcoming endocrine resistance. Breast Cancer Res Treat, 2005, 93 (suppl 1), S11-18.

[25.] Gradishar W, Chia S, Piccart M et al, on behalf of the EFECT writing committee: Fulvestrant versus exemestane following prior non-steroidal aromatase inhibitor therapy: first results from EFECT, a randomized, phase III trial in postmenopausal women with advanced breast cancer, San Antonio Breast Cancer Symposium, 2006, Abstr. 12.

[26.] Chen D, Washbrook E, Sarwar N et al. Phosphorylation of human estrogen receptor alpha at serine 118 by two distinct signal transduction pathways revealed by phosphorylation-specific antisera. Oncogene, 2002, 21, 4921-4931.

[27.] Kato S, Endoh H, Masuhiro Y et al. Activation of the estrogen receptor through phosphorylation by mitogen-activated protein kinase. Science, 1995, 270, 1491-1494.

[28.] Campbell RA, Bhat-Nakshatri P, Patel NM et al. Phosphatidylinositol 3-kinase/AKT-mediated activation of estrogen receptor alpha: a new model for anti-estrogen resistance. J Biol Chem, 2001, 276, 9817-9824.

[29.] Ali S and Coombes RC. Endocrine-responsive breast cancer and strategies for combating resistance. Nat Rev Cancer, 2002, 2, 101-112.

[30.] Dowsett M, Ebbs SR, Dixon JM et al. Biomarker changes during neoadjuvant anastrozole, tamoxifen, or the combination: influence of hormonal status and HER-2 in breast cancer-a study from the IMPACT trialists. J Clin Oncol, 2005, 23, 2477-2492.

[31.] Smith IE, Dowsett M, Ebbs SR et al. Neoadjuvant treatment of postmenopausal breast cancer with anastrozole, tamoxifen, or both in combination: the Immediate Preoperative Anastrozole, Tamoxifen, or Combined with Tamoxifen (IMPACT) multicenter double-blind randomized trial. J Clin Oncol, 2005, 23, 5108-5116.

[32.] Bautista S, Valles H, Walker RL et al. In breast cancer, amplification of the steroid receptor coactivator gene AIB1 is correlated with estrogen and progesterone receptor positivity. Clin Cancer Res, 1998, 4, 2925-2929.

[33.] Osborne CK, Bardou V, Hopp TA et al. Role of the estrogen receptor coactivator AIB1 (SRC-3) and HER-2/neu in tamoxifen resistance in breast cancer. J Natl Cancer Inst, 2003, 95, 353-361.

[34.] Kirkegaard T, McGlynn LM, Campbell FM et al. Amplified in breast cancer 1 in human epidermal growth factor receptor-positive tumors of tamoxifen-treated breast cancer patients. Clin Cancer Res, 2007, 13, 1405-1411.

[35.] Lavinsky RM, Jepsen K, Heinzel T et al. Diverse signaling pathways modulate nuclear receptor recruitment of N-CoR and SMRT complexes. Proc Natl Acad Sci U S A, 1998, 95, 2920-2925.

[36.] Zwijsen RM, Buckle RS, Hijmans EM et al. Ligand-independent recruitment of steroid receptor coactivators to estrogen receptor by cyclin D1. Genes Dev, 1998, 12, 3488-3498.

[37.] Kenny FS, Hui R, Musgrove EA et al. Overexpression of cyclin D1 messenger RNA predicts for poor prognosis in estrogen receptor-positive breast cancer. Clin Cancer Res, 1999, 5, 2069-2076.

[38.] Han S, Park K, Bae BN et al. Cyclin D1 expression and patient outcome after tamoxifen therapy in estrogen receptor positive metastatic breast cancer. Oncol Rep, 2003, 10, 141-144.

[39.] Johnston SR, Lu B, Scott GK et al. Increased activator protein-1 DNA binding and c-Jun NH2-terminal kinase activity in human breast tumors with acquired tamoxifen resistance. Clin Cancer Res, 1999, 5, 251-256.

[40.] Webb P, Lopez GN, Uht RM et al. Tamoxifen activation of the estrogen receptor/AP-1 pathway: potential origin for the cell-specific estrogen-like effects of antiestrogens. Mol Endocrinol, 1995, 9, 443-456.

[41.] Chung YL, Sheu ML, Yang SC et al. Resistance to tamoxifen-induced apoptosis is associated with direct interaction between Her2/neu and cell membrane estrogen receptor in breast cancer. Int J Cancer, 2002, 97, 306-312.

[42.] Osborne CK, Shou J, Massarweh S et al. Crosstalk between estrogen receptor and growth factor receptor pathways as a cause for endocrine therapy resistance in breast cancer. Clin Cancer Res, 2005, 11, 865s-870s.

[43.] Johnston SR. Combinations of endocrine and biological agents: present status of therapeutic and presurgical investigations. Clin Cancer Res, 2005, 11, 889s-899s.

[44.] Leary A and Dowsett M. Combination therapy with aromatase inhibitors: the next era of breast cancer treatment? Br J Cancer, 2006, 95, 661-666.

[45.] Giacinti L, Claudio PP, Lopez M et al. Epigenetic information and estrogen receptor alpha expression in breast cancer. Oncologist, 2006, 11, 1-8.

[46.] Yang X, Phillips DL, Ferguson AT et al. Synergistic activation of functional estrogen receptor (ER)-alpha by DNA methyltransferase and histone deacetylase inhibition in human ER-alpha-negative breast cancer cells. Cancer Res, 2001, 61, 7025-7029.

[47.] Ferguson AT, Lapidus RG, Baylin SB et al. Demethylation of the estrogen receptor gene in estrogen receptor-negative breast cancer cells can reactivate estrogen receptor gene expression. Cancer Res, 1995, 55, 2279-2283.

[48.] Zhang QX, Borg A, Wolf DM et al. An estrogen receptor mutant with strong hormone-independent activity from a metastatic breast cancer. Cancer Res, 1997, 57, 1244-1249.

[49.] Herynk MH and Fuqua SA. Estrogen receptor mutations in human disease. Endocr Rev, 2004, 25, 869-898.

[50.] Johnston SR, Haynes BP, Smith IE et al. Acquired tamoxifen resistance in human breast cancer and reduced intra-tumoral drug concentration. Lancet, 1993, 342, 1521-1522.

[51.] Dowsett M and Haynes BP. Hormonal effects of aromatase inhibitors: focus on premenopausal effects and interaction with tamoxifen. J Steroid Biochem Mol Biol, 2003, 86, 255-263.

[52.] Goetz MP, Rae JM, Suman VJ et al. Pharmacogenetics of tamoxifen biotransformation is associated with clinical outcomes of efficacy and hot flashes. J Clin Oncol, 2005, 23, 9312-9318.

[53.] Goetz MP, Knox SK, Suman VJ et al. The impact of cytochrome P450 2D6 metabolism in women receiving adjuvant tamoxifen. Breast Cancer Res Treat, 2007, 101, 113-121.

[54.] Chetrite GS, Cortes-Prieto J, Philippe JC et al. Comparison of estrogen concentrations, estrone sulfatase and aromatase activities in normal, and in cancerous, human breast tissues. J Steroid Biochem Mol Biol, 2000, 72, 23-27.

[55.] Miller WR, Anderson TJ and Jack WJ. Relationship between tumour aromatase activity, tumour characteristics and response to therapy. J Steroid Biochem Mol Biol, 1990, 37, 1055-1059.

[56.] Lin Z RS, Cotterill H, Rademaker A et al. Messenger RNA levels of estrogen-related genes in malignant breast tumor tissue predict responsiveness to aromatase inhibitors. San Antonio Breast Cancer Symposium, 2006, Abstr. 14.

[57.] Dunning AM, Dowsett M, Healey CS et al. Polymorphisms associated with circulating sex hormone levels in postmenopausal women. J Natl Cancer Inst, 2004, 96, 936-945.

[58.] Ma CX, Adjei AA, Salavaggione OE et al. Human aromatase: gene resequencing and functional genomics. Cancer Res, 2005, 65, 11071-11082.

[59.] Mita M, de Bono J, Mita A et al. A phase II and biologic correlative study investigating anastrozole in combination with gefinitib in post menopausal patients with estrogen receptor positive metasatic breast carcinoma who have previously failed hormonal therapy. Breast Cancer Res Treat, 2005, 94 (suppl S1), Abstr. 1117.

[60.] Dowsett M, Smith I, Skene A et al.; on behalf of the Study 0223 Trialists. Biological and clinical outcomes from a phase II placebo-controlled neoadjuvant study of anastrozole alone or with gefinitib in post menopausal women with ER/PgR+ breast cancer. Proc ASCO, 2006, 25, Abstr. 515.

[61.] Polychronis A, Sinnett HD, Hadjiminas D et al. Preoperative gefinitib versus gefinitib and anastrozole in post menopausal patients with oestrogen-receptor positive and epidermal-growth-factor-receptor-positive primary breast cancer; a double- blind placebo-controlled phase II randomised trial. Lancet Oncol, 2005, 6, 383-391.

[62.] Marcom PK, Isaacs C, Harris L et al. The combination of letrozole and trastuzumab as first or second-line biological therapy produces durable responses in a subset of HER2 positive and ER positive advanced breast cancers. Breast Cancer Res Treat, 2006, Epub ahead of print.

[63.] Mackey JR, Kaufman B, Clemens M et al. Trastuzumab prolongs progression-free survival in hormone-dependent and HER2-positive metastatic breast cancer. Breast Cancer Res Treat, 2007, 100 (suppl S1), Abstr. 3.

[64.] Awada AC, Cardoso F, Fontaine C et al. A phase Ib study of the mTOR inhibitor RAD001 (everolimus) in combination with letrozole (femara), investigating the safety and pharmokinetics in patients with advanced breast cancer stable or slowly progressing on letrozole. Breast Cancer Res Treat, 2004, 88 (suppl 1), Abstr. 6043.

[65.] Baselga J, Roche H, Fumoleau P et al. Treatment of postmenopausal women with locally advanced or metastatic breast cancer with letrozole alone or in combination with temsirolimus: a randomised, 3-arm, phase 2 study. Breast Cancer Res Treat, 2005, 94 (suppl 1), Abstr. 1068.

[66.] Chow LW, Shun Y, Jassem J et al. Phase 3 study of temsirolimus with letrozole or letrozole alone in postmenopausal women with locally advanced or metastatic breast cancer. Breast Cancer Res Treat, 2006, 100 (suppl S1), Abstr. 6091.

Alexandra Leary and Stephen Johnston

Royal Marsden Hospital, London, UK

Correspondence to: Alexandra Leary, Department of Medicine, Royal Marsden Hospital, Fulham Road, London, SW3 6JJ, UK (email:
Table 1: Postulated mechanisms of endocrine resistance and potential
avenues for therapy

Biological mechanism Therapeutic options / comments

Growth factor driven

Downregulation of ER expression Sequential growth factor--ER-
 and/or function targeted therapy
Switch to ER-independent Non-ER-targeted therapy
Enhanced genomic and non-genomic Dual ER and growth factor
 functions of ER targeting


ER silencing due to promoter Demethylating agents or HDAC
 hypermethylation inhibitors
E2 hypersensitivity Continued ED plus ER downregulator
 or growth factor inhibitor
ER mutations Rare
Altered tamoxifen metabolism and Higher tamoxifen doses in selected
 cellular clearance patients?
Aromatase expression and function Relevance unknown

ER, oestrogen receptor; E2, oestradiol; HDAC, histone deacetylase; ED,
oestrogen deprivation.

Table 2: Combinations of endocrine therapies with inhibitors of
membrane receptors or downstream effectors: selected completed trials
(data from [59]).

Clinical Clinical
setting Phase Intervention endpoints Reference

Combinations with EGFR inhibitor gefitinib (GEF)

MBC II ANA + GEF PR=0 SD=0 [59]
Endocrine ORR=61% vs [60]
 resistant 48%, P=0.067
Neoadjuvant II RCT ANA vs ANA + ORR=50% vs 54%
 PBC GEF x 16 [61]
Preoperative II RT GEF vs GEF +
EGFR+ only x 4-6

Combinations with trastuzumab (TRAS)

HER2+ MBC II TRAS + LET PR=26% SD=26% [62]
HER2 MBC (all III RCT ANA vs ANA + PFS=2.4 mo vs [63]
 pts were IHC TRAS 4.8 mo,
 3+ or FISH+) P=0.0016
 OS=23.9 mo vs
 28.5 mo,

mTor inhibitors: everolimus (EVE) or temsirolimus (TEM)

MBC Ib n=9 LET+EVE 1, 5 No grade 3/4 [64]
stable or or 10 mg toxicities
 slowly daily at 5 mg
 progressing (6 pts) or
 on letrozole 10 mg (3
MBC II n=92 Let vs LET+TEM ORR=45% vs 33% [65]
 10 mg daily vs 40%
 vs LET+TEM PFS=11.6 mo
 30 mg vs 11.5 mo
 intermittent vs 13.2 mo
MBC III RCT LET vs ORR=24% vs 24% [66]
 n=992 letrozole+ SD=19% vs
 TEM 16% PFS=9.2
 intermittent mo vs 9.2 mo

EGFR, epidermal growth factor receptor; MBC, metastatic breast cancer;
PBC, primary breast cancer; RCT, randomised controlled trial; RT,
randomised trial; PR, partial response; SD, stable disease; ORR,
objective response rate (PR+CR); ANA, anastrozole; LET, letrozole;
mTOR, mammalian target of rapamycin; PFS, progression-free survival;
OS, overall survival; vs, versus; +, positive; mo, months; pts,
patients; IHC, immunohistochemistry; FISH, fluorescence in situ

Table 3. Endocrine combinations: selected ongoing or recently closed

Clinical Trial Biological protocol
setting phase Intervention endpoints number

Endocrine combinations

MBC III FUL vs EXEM vs Serum hormone 00253422
Previously FUL+ANA levels
with AI

MBC III ANA vs ANA + Not available 00256698
1st line FUL

With EGFR inhibitors: gefitinib (GEF) or erlotinib (ERL)

Neoadjuvant II GEF x 2 wks Tumour biomarker 00206492
PBC followed by analysis at
(HER2+ only) GEF+TAM weeks 1, 2 and
 6, and surgery

Neoadjuvant II ANA + FUL + GEF Tumour biomarker 00206414
PBC analysis pre-

MBC II RCT TAM [+ or -] Correlate RR to 00069290
 GEF HER2/A1B1 status
 and other

MBC II RCT ANA [+ or -] biomarker study 00077025

MBC II RCT ANA [+ or -] None specified 00066378

MBC II FUL + GEF None specified 00234403

MBC II RT ANA + GEF vs Identify 00057941
 FUL + GEF biologic

MBC II LET + ERL Correlation of 00179296
 EGFR, HER2 and
 pERSer118 to

MBC II RT GEF [+ or -] Correlate early 00080743
TAM resistant TAM changes on PET
 and in plasma
 DNA to response

With trastuzumab (TRAS)

HER2+ MBC II TRAS Correlate 00238290
AI-resistant monotherapy benefit with
 until PD HER2 ECD at
 followed by baseline and
 TRAS + LET after treatment

HER2+ MBC III RCT ANA [+ or -] None specified 00022672

HER2+ MBC IV LET + TRAS None specified 00171847

With lapatinib (LAP)

MBC II LAP + TAM None specified 00118157
TAM resistant

MBC III RCT LET [+ or -] Biomarker and 00073528
 LAP genetic variant

With mTOR inhibitors: everolimus (EVE) and temsirolimus (TEM)

Neoadjuvant II RCT LET [+ or -] None specified 00107016

MBC II RCT LET [+ or -] None specified 00061971

Combination with HDAC inhibitor SAHA

MBC II TAM+SAHA Pre-and post 00365599
 treatment ER
 expression and

PBC, primary breast cancer; MBC, metastatic breast cancer; RCT,
randomised controlled trial; RT, randomised trial; RR, response rate;
ECD, extracellular domain: ANA, anastrozole; FUL, fulvestrant; EXEM,
exemestane; LET, letrozole; TAM, tamoxifen; SAHA, suberoylanalide
hydroxamic acid; PET, positron emission tomography.
COPYRIGHT 2007 Mediscript Ltd.
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2007 Gale, Cengage Learning. All rights reserved.

 Reader Opinion




Article Details
Printer friendly Cite/link Email Feedback
Author:Leary, Alexandra; Johnston, Stephen
Publication:Advances in Breast Cancer
Geographic Code:1USA
Date:Jun 1, 2007
Previous Article:The role of targeted therapy in breast cancer.
Next Article:HER2-directed therapies.

Related Articles
Protein may tie obesity to diabetes.
Bad news for boys: linking hypospadias and endocrine disruptors. (Science Selections).
Origins of obesity.
Berne and Levy Principles of Physiology, 4th ed.
Exploring the roots of diabetes: bisphenol a may promote insulin resistance.
Endocrine physiology, 2d ed.
Endocrine disruptors.
The role of targeted therapy in breast cancer.

Terms of use | Copyright © 2014 Farlex, Inc. | Feedback | For webmasters