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Relationship: 2612
Title
Increase activation, Nuclear factor kappa B (NF-kB) leads to Antagonism, Estrogen receptor
Upstream event
Downstream event
Key Event Relationship Overview
AOPs Referencing Relationship
AOP Name | Adjacency | Weight of Evidence | Quantitative Understanding | Point of Contact | Author Status | OECD Status |
---|---|---|---|---|---|---|
DNA damage and mutations leading to Metastatic Breast Cancer | adjacent | Moderate | Moderate | Agnes Aggy (send email) | Under development: Not open for comment. Do not cite |
Taxonomic Applicability
Sex Applicability
Life Stage Applicability
Key Event Relationship Description
Upstream event: Increased, NF kB activity
Downstream event: Estrogen receptor, Reduced
Evidence Collection Strategy
Evidence Supporting this KER
Activation NF-κB in breast cancer leads to loss of Estrogen Receptor (ER) expression and Human Epidermal Growth Fac- tor Receptor 2 (HER-2) overexpressed via epidermal growth factor receptor (EGFR) and Mitogen Activated Protein Kinase (MAPK) pathway (Laere et al.,2007). Indeed, the binding of epidermal growth factor (EGF) to its receptor (EGFR) activates NF-B, which most likely contributes to this transcription factor's increased activity in ER negative breast cancer cells (Shostak et al.,2011). Because of constitutive production of cytokines and growth factors, loss of ER function has been linked to constitutive NF-kB activity and hyperactive MAPK, resulting in aggressive, metastatic, hormone-resistant malignancies (Ali et al., 2002). Activation of the progesterone receptor can reduce DNA binding and transcriptional activity by inhibiting NF-B-driven gene expression (Kalkhoven et al., 1996). HER-2 stimulates NF-B via the conventional route, which includes IKK (Merkhofer et al., 2010).
Biological Plausibility
-NF-kB activation in breast cancer has been extensively documented in oestrogen receptor negative (ER) breast tumours and ER breast cancer cell lines, implying a significant inhibitory interaction between both signalling pathways (Biswas et al, 2000, 2001, 2004; Zhou et al, 2005). A rise in both NF-kB DNA-binding activity (Nakshatri et al, 1997) and expression of NF-kB target genes such IL8 coincides with a transition from oestrogen dependence to oestrogen independence in breast cancer, indicating inhibitory cross-talk. The fact that some breast tumours that are resistant to the tumoricidal action of anti-estrogens become sensitised to apoptosis and show a drop in NF-kB activity after treatment with oestrogen supports the inverse relationship between ER and NF-kB activity.
-This shows that oestrogen's proapoptotic actions in these tumours are mediated via NF-kB suppression.
Empirical Evidence
- In specific subclasses of human breast cancer cells and tumour tissue specimens, an enhanced level of activated NF-kB is found, primarily in erbB2-overexpressing ER-negative breast cancer (Biswas et al 2000;2003).
- Singh et al explored a variety of methods to inhibit NF-kB activation in ER-negative breast cancer cells and looked at the effects on cell proliferation, apoptosis, and tumour growth in xenografts(Singh et al.,2007). Several cell lines were utilised as representative cultured cell models for subclasses of human breast cancer, including ER negative and erbB2 positive (SKBr3 and MDA-MB453), ER negative and erbB1 positive (MDA-MB231), and ER positive and erbB1/erbB2 negative (MCF-7). IKK, the primary kinase in NF-nB activation, was disabled using a conditional dominant-negative gene construct and small-molecule inhibitors. Bortezomib, a proteasome inhibitor, was used to prevent NF-kB activation.
-The study compared the results of NF-kB activation patterns in ER-negative and erbB2-positive SKBr3 and MDA-MB453 cell lines as representative functional systems of this subclass of human breast cancer to the ER-positive and erbB2-negative MCF-7 cell line. Growing SKBr3 cells in deficient (minimum) media followed by supplementation with the particular mitogenic growth factor HRG (+HRG) allowed the degree of activated NF-kB to be experimentally controlled. This therapy increased IKK activity in as little as 15 minutes, while also increasing NF-nB DNA-binding activity and NF-nB–driven reporter gene expression. These findings imply that HRG-initiated signalling in erbB2-positive cells is mediated by IKK-induced NF-kB activation. MDA-MB453, a second erbB2-positive and ER-negative breast cancer cell line, showed similar results.
- In a prospective cohort study, Sampepajung et al used immunohistochemistry (IHC) to examine NF-B expression and intrinsic subtypes of breast cancer tissue and found a significant correlation between negative ER and overexpression of NF-B (p 0.05), with overexpression of NF-B being higher in negative ER (77.3 percent) compared to positive ER (47.4 percent )( Sampepajung et al., 2021)
- Laere et al suggested that activation of NF-kB in inflammatory breast cancer (IBC) is associated with loss of estrogen receptor (ER) expression, indicating a potential crosstalk between NF-kB and ER(Laere et al.,2007).
In this study, the activation of NF-kB in IBC and non-IBC cells was investigated in relation to ER and EGFR expression, ErbB2 expression, and MAPK hyperactivation.The expression of eight NF-kB target genes was associated with the expression of a qRT–PCR-based ER signature in tumours with and without transcriptionally active NF-kB. Hierarchical clustering was performed using a combined ER/NF-kB signature. MAPK hyperactivation was associated to tumour phenotype, ER and EGFR overexpression, and/or ErbB2 overexpression, according to a recently reported MAPK signature.
In breast tumours without transcriptionally active NF-kB, the expression of most ER-modulated genes was much higher. Furthermore, the expression of most ER-modulated genes was highly anticorrelated with that of most NF-kB target genes, demonstrating that ER and NF-kB activity are inversely related.
-The activation of the transcription factors of ER and NF-kB are inversely linked.
- Indra et al employed the prospective cohort approach to investigate 62 samples in an observational analysis.The positive and negative expression of NF-B, ER, and HER2 overexpression were among the data used in this investigation(Indra et al.,2021). The cases were separated into two groups: those who responded to neoadjuvant chemotherapy and those who did not. Negative NF-B expression (82.5%), positive HER2 status (85.7%), and negative ER status (85.7%) were all associated with a larger percentage of responding individuals (71.9 percent ).
-NF-B expression, ER status, and HER2 all had a substantial relationship with the response to anthracycline-based neoadjuvant chemotherapy for locally advanced breast cancer, with NF-B expression having the strongest link.
-A higher percentage of responding participants (71.9 percent) had a negative ER status, indicating that ER expression and chemotherapy response have a substantial relationship (p 0.05).
- This finding is consistent with alanalysis by Osako et al, of 103 individuals with locally advanced KPD. Neoadjuvant chemotherapy with anthracyclines and taxanes was given to the patients. The pCR chemotherapy results were significantly correlated with negative ER and PR expression results (Osako et al.,2012).
The expression of NF-B, HER2, and ER status has a strong association with chemotherapy response, according to these findings. Multivariate analysis of the specific association between NF-B expression and chemotherapeutic response revealed that NF-B expression and HER2 status were both related with chemotherapy response , whereas ER status had no such relationship.
- Sarkar et al assessed NF-B expression in breast cancer tissue and fibroadenoma tissue as test samples and controls, respectively. The Western Blot Technique was used to measure the p65 protein from the NF-kB superfamily of transcription factors. Immunohistochemistry was used to determine the levels of ER, PR, and HER-2/neu(Sarkar et al., 2013).Large tumour size (5 cm), high grade tumours, negative ER, negative PR, and positive HER-2/neu are all related with -NF-B/p65.
-NF-B activation was shown to be more common in ER-negative tumours (81.8%) than in ER-positive cancers (38.5%), a statistically significant difference.
-NF-B expression is linked to ER negative status and is also linked to a higher NPI value, which indicates a poor prognosis.
- NF-kB activity is elevated in hormone-independent and ER-negative breast tumors , and hyperactiva- tion of MAPK leads to enhanced NF-kB activity through induction of autocrine factors such as HB- EGF (Norris et l.,1999;Pearson et l.,2001;McCarthy et l.,1995;Troppamair et l.,1998). NF-kB activity is elevated in MCF-7 breast cancer cells with elevated MAPK activity(Holloway et l.,2004).
-NF-kB activity is about 5-fold higher than parental MCF-7 in all of our model cell lines. This elevated NF-kB activity is attributable to hyperactivation of MAPK because NF-kB activity is returned to normal levels (basal levels in co-MCF7 cells) by dnERKs 1 and 2 .
Uncertainties and Inconsistencies
No specific uncertainties and inconsistencies reported to the best of our knowledge.
Known modulating factors
Estradiol has been shown to decrease transcriptional activity and expression of NF-kB in a variety of experimental models (Biswas et al., 2005;Lobanova et al.,2007). Estrogen treatment of MCF-7 or MCF-7/H cells resulted in a significant suppression of NF-kB activity in both cell lines, according to research. The antiestrogen tamoxifen boosted NF-kB activity in the cells, indicating that ER plays a key role in NF-kB down-regulation in both parent and hypoxia-tolerant cells.
-MCF-7/T2H cells were discovered to have a partial tolerance to acute cobalt chloride-induced hypoxia while maintaining their estrogen-independent phenotype. In contrast to the MCF-7/H subline, MCF-7/T2H cells had a non-affected baseline NF-kB level, indicating that estrogens are responsible for NF-kB downregulation (Scherbakov et al., 2009).
Quantitative Understanding of the Linkage
Method/ measurement reference |
Reliability |
Strength of evidence |
Assay fit for purpose |
Repeatability/ reproducibility |
Direct measure |
|
Human cell line |
qPCR, western blotting, immunoprecipitation, immunofluorescent microscopy, Luciferase reporter assay EMSA, IHC,Cell viability assay |
Yes |
Strong |
Yes |
Yes |
Yes |
Humans |
qRT-PCR, immunohistochemistry |
Yes |
Strong |
Yes |
Yes |
Yes |
Mouse(A1) |
EMSA,Autoradiography Immunofluorescent microscopy, Westernblotting |
Yes |
Strong |
Yes |
Yes |
Yes |
Response-response Relationship
Differential Sensitivity of ER α and ERβ Cells to the NF-kB Inhibitor Go6976. A differential sensitivity to Go6976 by ER α and ERβ breast cancer cells was observed (Holloway et al.,2004). The ER α cells were more sensitive and less viable after treatment with this NF-kB inhibitor. The IC50 (50% killing) by Go6976 was 1 mM for Era of MDA-MB435 and MDA-MB231 breast cancer cells, whereas it was greater than 10 mM for ERa of MCF-7 and T47D or the normal mammary epithelial H16N cells . At 10 mM Go6976, about 80% of the ERa cells were killed, whereas only 15–30% of ERa and normal H16N cells were sensitive to this compound. The relative resistance of the H16N normal human mammary cells indicates a possible high therapeutic index of Go6976 against ERa cancer cells.
This observation is consistent with the previously observed role of NF-kB as an antiapoptotic agent. FACS analysis demonstrated accumulation of sub-G1 population (67%) in Go6976- treated (48 h at 1 mM) ERa vs. only 10–15% in ERa cells, indicating enhanced apoptotic cell death preferentially of ERa cells caused by this low molecular weight compound.
Time-scale
Key events connected by this KER occur within hours of exposure.
Known Feedforward/Feedback loops influencing this KER
- Multiple pathways are implicated in the crosstalk between NF-KB and ER. Through many mechanisms, including collaboration with FOXA1 to strengthen latent ER-binding sites and trigger translation of their synergistic genes, NF-KB directly interacts with the DNA-binding activity of ER (Franco et al. 2015). Furthermore, NF-KB affects ER via interacting with its ER co-activator or co-repressor, which changes ER transcriptional activity (Park et al. 2005). Similar to ER, NF-KB has been reported to have a role as a downstream effector for the growth factor pathway, which is recognized to be involved in both ligand-dependent and non-ligand-dependent ER activation, leading to resistance to a wide range of anti- oestrogen drugs (Zhou et al. 2005a, Sas et al. 2012, Frasor et al. 2015).
-NF-KB is also involved in the anti-apoptotic pathway and immune surveillance systems, both of which have been linked to endocrine resistance (Hu et al. 2015; Lim et al. 2016). Furthermore, NF-KB inhibition of ER activity has been observed. The zinc finger repressor B-lymphocyte-induced maturation protein (BLIMP1), which can bind to the ER promoter area and restrict ER transcription, is triggered by the NFB subunit RelB. (Wang et al. 2009). Increasing data suggests that NF-KB plays an important role in the complexities of the endocrine resistance environment in breast cancer.
-NF-KB and ERS1 mutations in breast cancer patients who are resistant to endocrine therapy
TNF needs NF-KB and FOXA1 to change the breast cancer cell transcriptome by modulating latent ER-binding sites.
Domain of Applicability
KER has been observed in humans and animals irrespective of the gender and life stage.
References
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Kalkhoven, E., Wissink, S., van der Saag, P. T., & van der Burg, B. (1996). Negative Interaction between the RelA (p65) Subunit of NF-κB and the Progesterone Receptor (∗). Journal of Biological Chemistry, 271(11), 6217-6224.
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