Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Aug 5;10(8):1986.
doi: 10.3390/cells10081986.

Neuroglobin: A New Possible Marker of Estrogen-Responsive Breast Cancer

Virginia Solar Fernandez  1 , Marco Fiocchetti  1 , Manuela Cipolletti  1 , Marco Segatto  2 , Paolo Cercola  3 , Annalisa Massari  3 , Sabrina Ghinassi  3 , Francesco Cavaliere  3 , Maria Marino  1
Affiliations

Neuroglobin: A New Possible Marker of Estrogen-Responsive Breast Cancer

Virginia Solar Fernandez et al. Cells. .

Abstract

The expression of the α-subtype of Estrogen Receptor (ERα) characterizes most breast cancers (more than 75%), for which endocrine therapy is the mainstay for their treatment. However, a high percentage of ERα+ breast cancers are de novo or acquired resistance to endocrine therapy, and the definition of new targets for improving therapeutic interventions and the prediction of treatment response is demanding. Our previous data identified the ERα/AKT/neuroglobin (NGB) pathway as a common pro-survival process activated in different ERα breast cancer cell lines. However, no in vivo association between the globin and the malignity of breast cancer has yet been done. Here, we evaluated the levels and localization of NGB in ERα+ breast ductal carcinoma tissue of different grades derived from pre-and post-menopausal patients. The results indicate a strong association between NGB accumulation, ERα, AKT activation, and the G3 grade, while no association with the menopausal state has been evidenced. Analyses of the data set (e.g., GOBO) strengthen the idea that NGB accumulation could be linked to tumor cell aggressiveness (high grade) and resistance to treatment. These data support the view that NGB accumulation, mainly related to ER expression and tumor grade, represents a compensatory process, which allows cancer cells to survive in an unfavorable environment.

Keywords: AKT; breast cancer survival; ductal infiltrating adenocarcinoma; estrogen receptor α; neuroglobin; tumor microenvironment.

PubMed Disclaimer

Conflict of interest statement

The authors declare that they have no conflict of interest.

Figures

Figure 1
Figure 1
NGB levels in breast cancer tissue. Representative Western blot images of NGB protein levels (A) in two G2-grade post-menopausal specimens of breast cancer (T) and their normal counterpart (N). The amount of protein was normalized by comparison with vinculin levels. A total of 5 ng of recombinant NGB was used as protein standard. Data are the mean ± SD of 26 tissues. * p < 0.001 was calculated with Student’s t-test with respect to normal tissues. (B) Representative immunohistochemical staining of NGB (mouse monoclonal Clone 6G1.1 anti-NGB MERCK) in 1 out of 10 breast G2-grade post-menopausal specimens of breast cancer (T) and normal counterpart (N). The scale bar is 100 μm/cm. The black square refers to the below-reported optical magnification.
Figure 2
Figure 2
Association between NGB levels and estrogen receptors in breast cancer tissue. Representative Western blot images of NGB, pERα, ERα, ERβ, GPER, Bcl-2 protein levels (A) and AKT levels and activation (B) in G2-grade post- (n = 26) and pre-menopausal (n = 12) specimens of breast cancer (T) and their normal counterpart (N). The amount of protein was normalized by comparison with vinculin levels. Data are the mean ± SD. p < 0.001 was calculated with Student’s t-test with respect to the normal or tumoral (e.g., GPER) counterpart (*) and vs. the same parameter on the post-menopausal sample (°).
Figure 3
Figure 3
NGB protein levels in tumor samples of breast cancer patients. Representative Western blot images (upper panel) of tissue levels of NGB from G2 and G3 samples and correspondent densitometric analysis of all analyzed tissues (n = 38, n = 15, respectively) (bottom panel). The amount of protein was normalized by comparison with vinculin levels. Data are means ± SD. p < 0.01 was determined with ANOVA followed by Bonferroni post-test vs. G2 (*) samples.
Figure 4
Figure 4
NGB localization in breast tissues. (A) Representative Western blot image (left panel) of NGB levels in whole tissue (Tissue lysate) and in subcellular (Mitochondria-Mito, Cytosol-Cyto) fractions of G2 and G3 samples and densitometric analysis (right panel) of NGB levels in subcellular fractions among all available tissues (G2 n = 38, G3 n = 15, respectively). The amount of protein was normalized by comparison with the cytosolic marker protein phosphatase 2A (PP2A) and the mitochondrial marker tumor necrosis factor-associated protein 1 (TRAP-1). Data are means ± SD. p < 0.01 was determined with ANOVA followed by Bonferroni post-test vs. G2 (*) samples and vs. correspondent mitochondrial fraction (°). (B) Representative image shows positive NGB staining (mouse monoclonal Clone 6G1.1 anti-NGB MERCK) in the epithelial cells of the tumoral section in a G2 ERα+ breast cancer tissue sample (total n = 10). The scale bar is 100 μm/cm. The black square refers to image detail reported on the right as a digital magnification.
Figure 5
Figure 5
Data set analysis of survival in patients with different NGB expressions. Kaplan–Meier plots of high (red line) or low (gray line) NGB-expressing tumor samples (ID = 58157) available in the GOBO (Gene expression-based Outcome for Breast cancer Online). The data have been selected from ERα+ samples based on tumor grade (lowest at left, highest at right). The overall (upper panels) and the relapse-free (bottom panels) survival have been selected as outcomes.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Sung H., Ferlay J., Siegel R.L., Laversanne M., Soerjomataram I., Jemal A., Bray F. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA A Cancer J. Clin. 2021;71:209–249. doi: 10.3322/caac.21660. - DOI - PubMed
    1. DeSantis C.E., Ma J., Gaudet M.M., Newman L.A., Miller K.D., Sauer A.G., Jemal A., Siegel R.L. Breast Cancer Statistics, 2019. CA A Cancer J. Clin. 2019;69:438–451. doi: 10.3322/caac.21583. - DOI - PubMed
    1. Acconcia F., Marino M. The Effects of 17b-Estradiol in Cancer Are Mediated by Estrogen Receptor Signaling at the Plasma Membrane. Front. Physiol. 2011;2:30. doi: 10.3389/fphys.2011.00030. - DOI - PMC - PubMed
    1. Skandalis S.S., Afratis N., Smirlaki G., Nikitovic D., Theocharis A.D., Tzanakakis G.N., Karamanos N.K. Cross-Talk between Estradiol Receptor and EGFR/IGF-IR Signaling Pathways in Estrogen-Responsive Breast Cancers: Focus on the Role and Impact of Proteoglycans. Matrix Biol. 2014;35:182–193. doi: 10.1016/j.matbio.2013.09.002. - DOI - PubMed
    1. Vuong D., Simpson P.T., Green B., Cummings M.C., Lakhani S.R. Molecular Classification of Breast Cancer. Virchows Arch. 2014;465:1–14. doi: 10.1007/s00428-014-1593-7. - DOI - PubMed

Publication types

MeSH terms