Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

α-catenin acts as a tumour suppressor in E-cadherin-negative basal-like breast cancer by inhibiting NF-κB signalling

Nature Cell Biology volume 16, pages 245–254 (2014)Cite this article

Subjects

Abstract

Basal-like breast cancer is a highly aggressive tumour subtype associated with poor prognosis. Aberrant activation of NF-κB signalling is frequently found in triple-negative basal-like breast cancer cells, but the cause of this activation has remained elusive.Here we report that α-catenin functions as a tumour suppressor in E-cadherin-negative basal-like breast cancer cells by inhibiting NF-κB signalling. Mechanistically, α-catenin interacts with the IκBα protein, and stabilizes IκBα by inhibiting its ubiquitylation and its association with the proteasome. This stabilization in turn prevents nuclear localization of RelA and p50, leading to decreased expression of TNF-α, IL-8 and RelB. In human breast cancer, CTNNA1 expression is specifically downregulated in the basal-like subtype, correlates with clinical outcome and inversely correlates with TNF and RELB expression. Taken together, these results uncover a previously undescribed mechanism by which the NF-κB pathway is activated in E-cadherin-negative basal-like breast cancer.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: α-catenin inhibits proliferation and colony formation of basal-like breast cancer cells.
Figure 2: α-catenin inhibits NF-κB signalling in basal-like breast cancer cells.
Figure 3: α-catenin stabilizes IκBα protein by inhibiting IκBα ubiquitylation and abrogating IκBα interaction with the proteasome.
Figure 4: α-catenin inhibits RelA–p50 nuclear localization and downregulates RelB.
Figure 5: Loss of α-catenin promotes tumour growth in basal-like breast cancer cells by activating NF-κB signalling.
Figure 6: α-catenin inhibits tumorigenesis and is downregulated in human basal-like breast cancer.

Similar content being viewed by others

References

  1. Di Cosimo, S. & Baselga, J. Management of breast cancer with targeted agents: importance of heterogeneity. [corrected]. Nat. Rev. Clin. Oncol. 7, 139–147 (2010).

    Article  PubMed  Google Scholar 

  2. Perou, C. M. et al. Molecular portraits of human breast tumours. Nature 406, 747–752 (2000).

    Article  CAS  PubMed  Google Scholar 

  3. Sorlie, T. et al. Repeated observation of breast tumor subtypes in independent gene expression data sets. Proc. Natl Acad. Sci. USA 100, 8418–8423 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Prat, A. & Perou, C. M. Deconstructing the molecular portraits of breast cancer. Mol. Oncol. 5, 5–23 (2011).

    Article  CAS  PubMed  Google Scholar 

  5. Rakha, E. A., El-Sayed, M. E., Reis-Filho, J. & Ellis, I. O. Patho-biological aspects of basal-like breast cancer. Breast Cancer Res. Treat. 113, 411–422 (2009).

    Article  PubMed  Google Scholar 

  6. Perou, C. M. Molecular stratification of triple-negative breast cancers. Oncologist 16 (Suppl 1), 61–70 (2011).

    Article  Google Scholar 

  7. Osborne, C. K. Tamoxifen in the treatment of breast cancer. New Engl. J. Med. 339, 1609–1618 (1998).

    Article  CAS  PubMed  Google Scholar 

  8. Slamon, D. J. et al. Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science 244, 707–712 (1989).

    Article  CAS  PubMed  Google Scholar 

  9. Kobielak, A. & Fuchs, E. α-catenin: at the junction of intercellular adhesion and actin dynamics. Nat. Rev. Mol. Cell Biol. 5, 614–625 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. The Cancer Genome Atlas Network, Comprehensive molecular portraits of human breast tumours. Nature 490, 61–70 (2012).

    Article  PubMed Central  Google Scholar 

  11. Onder, T. T. et al. Loss of E-cadherin promotes metastasis via multiple downstream transcriptional pathways. Cancer Res. 68, 3645–3654 (2008).

    Article  CAS  PubMed  Google Scholar 

  12. Derksen, P. W. et al. Somatic inactivation of E-cadherin and p53 in mice leads to metastatic lobular mammary carcinoma through induction of anoikis resistance and angiogenesis. Cancer Cell 10, 437–449 (2006).

    Article  CAS  PubMed  Google Scholar 

  13. Fodde, R. & Brabletz, T. Wnt/β-catenin signaling in cancer stemness and malignant behavior. Curr. Opin. Cell Biol. 19, 150–158 (2007).

    Article  CAS  PubMed  Google Scholar 

  14. Liu, T. X. et al. Chromosome 5q deletion and epigenetic suppression of the gene encoding α-catenin (CTNNA1) in myeloid cell transformation. Nat. Med. 13, 78–83 (2007).

    Article  PubMed  Google Scholar 

  15. Ji, H., Wang, J., Fang, B., Fang, X. & Lu, Z. α-Catenin inhibits glioma cell migration, invasion, and proliferation by suppression of β-catenin transactivation. J. Neurooncol. 103, 445–451 (2011).

    Article  CAS  PubMed  Google Scholar 

  16. Schlegelmilch, K. et al. Yap1 acts downstream of α-catenin to control epidermal proliferation. Cell 144, 782–795 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Silvis, M. R. et al. α-catenin is a tumor suppressor that controls cell accumulation by regulating the localization and activity of the transcriptional coactivator Yap1. Sci. Signal. 4, ra33 (2011).

    Article  PubMed  PubMed Central  Google Scholar 

  18. Bajpai, S., Feng, Y., Krishnamurthy, R., Longmore, G. D. & Wirtz, D. Loss of α-catenin decreases the strength of single E-cadherin bonds between human cancer cells. J. Biol. Chem. 284, 18252–18259 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Yamaguchi, N. et al. Constitutive activation of nuclear factor-κB is preferentially involved in the proliferation of basal-like subtype breast cancer cell lines. Cancer Sci. 100, 1668–1674 (2009).

    Article  CAS  PubMed  Google Scholar 

  20. Baud, V. & Karin, M. Is NF-κB a good target for cancer therapy? Hopes and pitfalls. Nat. Rev. Drug Discov. 8, 33–40 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Pahl, H. L. Activators and target genes of Rel/NF-κB transcription factors. Oncogene 18, 6853–6866 (1999).

    Article  CAS  PubMed  Google Scholar 

  22. Hayden, M. S. & Ghosh, S. Signaling to NF-κB. Genes Dev. 18, 2195–2224 (2004).

    Article  CAS  PubMed  Google Scholar 

  23. Baltimore, D. Discovering NF-κB. Cold Spring Harb. Perspect. Biol. 1, a000026 (2009).

    Article  PubMed  PubMed Central  Google Scholar 

  24. Karin, M., Yamamoto, Y. & Wang, Q. M. The IKK NF-κB system: a treasure trove for drug development. Nat. Rev. Drug Discov. 3, 17–26 (2004).

    Article  CAS  PubMed  Google Scholar 

  25. Basseres, D. S. & Baldwin, A. S. Nuclear factor-κB and inhibitor of κB kinase pathways in oncogenic initiation and progression. Oncogene 25, 6817–6830 (2006).

    Article  CAS  PubMed  Google Scholar 

  26. Hollestelle, A. et al. Four human breast cancer cell lines with biallelic inactivating α-catenin gene mutations. Breast Cancer Res. Treat. 122, 125–133 (2010).

    Article  CAS  PubMed  Google Scholar 

  27. Collart, M. A., Baeuerle, P. & Vassalli, P. Regulation of tumor necrosis factor α transcription in macrophages: involvement of four κB-like motifs and of constitutive and inducible forms of NF-κB. Mol. Cell Biol. 10, 1498–1506 (1990).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Kunsch, C. & Rosen, C. A. NF-κB subunit-specific regulation of the interleukin-8 promoter. Mol. Cell Biol. 13, 6137–6146 (1993).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Vincenti, M. P., Coon, C. I. & Brinckerhoff, C. E. Nuclear factor κB/p50 activates an element in the distal matrix metalloproteinase 1 promoter in interleukin-1η-stimulated synovial fibroblasts. Arthritis Rheum. 41, 1987–1994 (1998).

    Article  CAS  PubMed  Google Scholar 

  30. Matluk, N., Rochira, J. A., Karaczyn, A., Adams, T. & Verdi, J. M. A role for NRAGE in NF-κB activation through the non-canonical BMP pathway. BMC Biol. 8, 7 (2010).

    Article  PubMed  PubMed Central  Google Scholar 

  31. Lien, W. H., Gelfand, V. I. & Vasioukhin, V. α-E-catenin binds to dynamitin and regulates dynactin-mediated intracellular traffic. J. Cell Biol. 183, 989–997 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Pickart, C. M. & Fushman, D. Polyubiquitin chains: polymeric protein signals. Curr. Opin. Chem. Biol. 8, 610–616 (2004).

    Article  CAS  PubMed  Google Scholar 

  33. Bren, G. D. et al. Transcription of the RelB gene is regulated by NF-κB. Oncogene 20, 7722–7733 (2001).

    Article  CAS  PubMed  Google Scholar 

  34. Gyorffy, B. et al. An online survival analysis tool to rapidly assess the effect of 22,277 genes on breast cancer prognosis using microarray data of 1,809 patients. Breast Cancer Res. Treat. 123, 725–731 (2010).

    Article  PubMed  Google Scholar 

  35. Ding, L. et al. Genome remodelling in a basal-like breast cancer metastasis and xenograft. Nature 464, 999–1005 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Kobielak, A. & Fuchs, E. Links between α-catenin, NF-κB, and squamous cell carcinoma in skin. Proc. Natl Acad. Sci. USA 103, 2322–2327 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Hu, Y. et al. Abnormal morphogenesis but intact IKK activation in mice lacking the IKKα subunit of IκB kinase. Science 284, 316–320 (1999).

    Article  CAS  PubMed  Google Scholar 

  38. Yan, J. et al. Inactivation of BAD by IKK inhibits TNFα-induced apoptosis independently of NF-κB activation. Cell 152, 304–315 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Ben-Neriah, Y. & Karin, M. Inflammation meets cancer, with NF-κB as the matchmaker. Nat. Immunol. 12, 715–723 (2011).

    Article  CAS  PubMed  Google Scholar 

  40. Gilmore, T. D. Introduction to NF-κB: players, pathways, perspectives. Oncogene 25, 6680–6684 (2006).

    Article  CAS  PubMed  Google Scholar 

  41. Karin, M., Cao, Y., Greten, F. R. & Li, Z. W. NF-κB in cancer: from innocent bystander to major culprit. Nat. Rev. Cancer 2, 301–310 (2002).

    Article  CAS  PubMed  Google Scholar 

  42. Pianetti, S., Arsura, M., Romieu-Mourez, R., Coffey, R. J. & Sonenshein, G. E. Her-2/neu overexpression induces NF-κB via a PI3-kinase/Akt pathway involving calpain-mediated degradation of IκB-α that can be inhibited by the tumor suppressor PTEN. Oncogene 20, 1287–1299 (2001).

    Article  CAS  PubMed  Google Scholar 

  43. Elenbaas, B. et al. Human breast cancer cells generated by oncogenic transformation of primary mammary epithelial cells. Genes Dev. 15, 50–65 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Stewart, S. A. et al. Lentivirus-delivered stable gene silencing by RNAi in primary cells. RNA 9, 493–501 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Yang, W. L. et al. The E3 ligase TRAF6 regulates Akt ubiquitination and activation. Science 325, 1134–1138 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Wang, W. et al. PTPN14 is required for the density-dependent control of YAP1. Genes Dev. 26, 1959–1971 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank S. Ethier (Medical University of South Carolina, USA) for the SUM159 cell line; J. Chen (The University of Texas MD Anderson Cancer Center, USA) for plasmids; the shRNA and ORFeome Core and the Histology Core at The University of Texas MD Anderson Cancer Center for technical assistance; J. Zhang and J. Chen for assistance with graphics; J. M. Rosen, X. Lin, W. Wang and members of the Ma laboratory for discussion; and H-l. Piao, C. Chu and A. Gelmis for editing the manuscript. This work is supported by US National Institutes of Health grants R00CA138572 (to L.M.) and R01CA166051 (to L.M.) and a Cancer Prevention and Research Institute of Texas Scholar Award R1004 (to L.M.).

Author information

Authors and Affiliations

  1. Department of Experimental Radiation Oncology, The University of Texas MD Anderson Cancer Center Houston Texas 77030 USA,

    Hai-long Piao, Min Wang & Li Ma

  2. Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center Houston Texas 77030 USA,

    Yuan Yuan & Han Liang

  3. Graduate Program in Structural and Computational Biology and Molecular Biophysics, Baylor College of Medicine Houston Texas 77030 USA,

    Yuan Yuan

  4. Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center Houston Texas 77030 USA,

    Yutong Sun

  5. Cancer Biology Program, Graduate School of Biomedical Sciences, The University of Texas Health Science Center at Houston Houston Texas 77030 USA,

    Li Ma

Authors
  1. Hai-long Piao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuan Yuan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Min Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yutong Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Han Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Li Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

L.M. conceived and supervised the project. H-l.P. designed, performed and analysed most of the experiments. Y.Y. and H.L. performed computational data analysis. M.W. performed immunohistochemical staining and provided animal care. Y.S. maintained shRNA and ORF clones and provided significant intellectual input. H-l.P. and L.M. wrote the manuscript with input from all other authors.

Corresponding author

Correspondence to Li Ma.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 α-catenin does not inhibit NF-κB signaling in E-cadherin-positive breast cancer cells.

(a) qPCR of CTNNA1, NFKBIA, RELB, IL8, TNF, SOD2 and BIRC3 in α-catenin-transduced MDA-MB-468 cells. n = 3 samples per group. (b) ELISA of TNFα and IL-8 secreted by α-catenin-transduced MDA-MB-468 cells. n = 3 wells per group. (c) Immunoblotting of α-catenin, RelB, IκBα, RelA, p105, p50 and HSP90 in α-catenin-transduced MDA-MB-468 cells. Data in (a) and (b) are the mean of biological replicates from a representative experiment, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t-test. The experiments were repeated three times. The source data can be found in Supplementary Table 4. Uncropped images of blots are shown in Supplementary Figure 7.

Supplementary Figure 2 α-catenin does not regulate IKK phosphorylation in MDA-MB-157 cells.

Endogenous IKKη was immunoprecipitated from mock-infected or α-catenin-transduced MDA-MB157 cells and immunoblotted with antibodies to p-IKK and IKKη. Cells were treated with TNFα (20 ng/ml). Uncropped images of blots are shown in Supplementary Figure 7.

Supplementary Figure 3 α-catenin does not regulate IκBα, RelA and RelB protein levels in luminal-like mammary cells.

(a) Immunoblotting of α-catenin, IκBα, RelA, RelB and HSP90 in T47D and MCF10A cells transduced with two independent α-catenin shRNAs. (b, c) Immunoblotting of α-catenin, IκBα, p-RelA, RelA and HSP90 in α-catenin shRNA-transduced T47D (b) and MCF10A (c) cells, with or without TNFα treatment (20 ng/ml). Uncropped images of blots are shown in Supplementary Figure 7.

Supplementary Figure 4 α-catenin does not modulate β-catenin in basal-like breast cancer cells.

(a) Immunoblotting of α-catenin and β-catenin in cytoplasmic and nuclear fractions of BT549 cells transduced with two independent α-catenin shRNAs. (b) Immunoblotting of α-catenin and β-catenin in cytoplasmic and nuclear fractions of α-catenin-transduced MDA-MB-157 cells. α-tubulin and lamin A were used as cytoplasmic and nuclear markers, respectively, in (a) and (b). (c) Luciferase assays of β-catenin activity in MDA-MB-157 cells transfected with the Fopflash or Topflash luciferase reporter alone or in combination with α-catenin or β-catenin, or both. The Topflash construct contains multiple TCF/LEF-binding sites in the promoter of a firefly luciferase reporter gene and the derived Fopflash construct contains mutated TCF/LEF binding sites. n = 3 wells per group. Data in (c) are the mean of biological replicates from a representative experiment, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t-test. The experiments were repeated three times. The source data can be found in Supplementary Table 4. Uncropped images of blots are shown in Supplementary Figure 7.

Supplementary Figure 5 α-catenin negatively correlates with NF-κB signaling components in human breast tumors.

(a) Box plots comparing CTNNA1 expression in normal breast tissues and in luminal A, luminal B and HER2+ breast tumors (n = 32, 13 and 4 patient samples, respectively). Statistical significance was determined by the Wilcoxon test. The boxes show the median and the interquartile range. The whiskers show the minimum and maximum. (b) Kaplan-Meier curves of relapse-free survival times of total breast cancer patients and patients with luminal A, luminal B or HER2+ breast cancer (n = 1370, 869 and 161 patient samples, respectively), stratified by CTNNA1expression levels. Statistical significance was determined by the log-rank test. (c) Scatterplots showing the inverse correlation of CTNNA1 with TNF (left panel) or RELB (right panel) expression in human breast tumors, based on the microarray data from TCGA. Statistical significance was determined by Spearman rank correlation test. Rs= Spearman rank correlation coefficient.

Supplementary Figure 6 α-catenin mechanisms of action.

(a) Cytoplasmic α-catenin and α-catenin localized to the cadherin-catenin complex regulate different pathways through different mechanisms. C: cytoplasm; N: nucleus; Ub: ubiquitin chain. (b) Immunoblotting of α-catenin, p-YAP, YAP and HSP90 in mock-infected and α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. Uncropped images of blots are shown in Supplementary Figure 7.

Supplementary information

Supplementary Information

Supplementary Information (PDF 969 kb)

Supplementary Table 1

Supplementary Information (XLSX 14 kb)

Supplementary Table 2

Supplementary Information (XLSX 13 kb)

Supplementary Table 3

Supplementary Information (XLSX 13 kb)

Supplementary Table 4

Supplementary Information (XLSX 36 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Piao, Hl., Yuan, Y., Wang, M. et al. α-catenin acts as a tumour suppressor in E-cadherin-negative basal-like breast cancer by inhibiting NF-κB signalling. Nat Cell Biol 16, 245–254 (2014). https://doi.org/10.1038/ncb2909

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb2909

This article is cited by

Search

Advanced search

Quick links

Nature Briefing: Cancer

Sign up for the Nature Briefing: Cancer newsletter — what matters in cancer research, free to your inbox weekly.

Get what matters in cancer research, free to your inbox weekly. Sign up for Nature Briefing: Cancer