Translational Therapeutics

HDAC5-mediated deacetylation and nuclear localisation of SOX9 is critical for tamoxifen resistance in breast cancer

Article metrics

Abstract

Background

Tamoxifen resistance remains a significant clinical challenge for the therapy of ER-positive breast cancer. It has been reported that the upregulation of transcription factor SOX9 in ER+ recurrent cancer is sufficient for tamoxifen resistance. However, the mechanisms underlying the regulation of SOX9 remain largely unknown.

Methods

The acetylation level of SOX9 was detected by immunoprecipitation and western blotting. The expressions of HDACs and SIRTs were evaluated by qRT-PCR. Cell growth was measured by performing MTT assay. ALDH-positive breast cancer stem cells were evaluated by flow cytometry. Interaction between HDAC5 and SOX9 was determined by immunoprecipitation assay.

Results

Deacetylation is required for SOX9 nuclear translocation in tamoxifen-resistant breast cancer cells. Furthermore, HDAC5 is the key deacetylase responsible for SOX9 deacetylation and subsequent nuclear translocation. In addition, the transcription factor C-MYC directly promotes the expression of HDAC5 in tamoxifen resistant breast cancer cells. For clinical relevance, high SOX9 and HDAC5 expression are associated with lower survival rates in breast cancer patients treated with tamoxifen.

Conclusions

This study reveals that HDAC5 regulated by C-MYC is essential for SOX9 deacetylation and nuclear localisation, which is critical for tamoxifen resistance. These results indicate a potential therapy strategy for ER+ breast cancer by targeting C-MYC/HDAC5/SOX9 axis.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

References

  1. 1.

    Ojo, D., Wei, F., Liu, Y., Wang, E., Zhang, H., Lin, X. et al. Factors promoting tamoxifen resistance in breast cancer via stimulating breast cancer stem cell expansion. Curr. Med. Chem. 22, 2360–2374 (2015).

  2. 2.

    Jeselsohn, R., Cornwell, M., Pun, M., Buchwalter, G., Nguyen, M., Bango, C. et al. Embryonic transcription factor SOX9 drives breast cancer endocrine resistance. Proc. Natl Acad. Sci. USA 114, E4482–E4491 (2017).

  3. 3.

    Abdelalim, E. M., Emara, M. M. & Kolatkar, P. R. The SOX transcription factors as key players in pluripotent stem cells. Stem. Cells Dev. 23, 2687–2699 (2014).

  4. 4.

    Lefebvre, V. & Dvir-Ginzberg, M. SOX9 and the many facets of its regulation in the chondrocyte lineage. Connect. Tissue Res. 58, 2–14 (2017).

  5. 5.

    Boiani, M. & Scholer, H. R. Regulatory networks in embryo-derived pluripotent stem cells. Nat. Rev. Mol. Cell. Biol. 6, 872–884 (2005).

  6. 6.

    Guo, W., Keckesova, Z., Donaher, J. L., Shibue, T., Tischler, V., Reinhardt, F. et al. Slug and Sox9 cooperatively determine the mammary stem cell state. Cell 148, 1015–1028 (2012).

  7. 7.

    Larsimont, J. C., Youssef, K. K., Sanchez-Danes, A., Sukumaran, V., Defrance, M., Delatte, B. et al. Sox9 controls self-renewal of oncogene targeted cells and links tumor initiation and invasion. Cell Stem Cell 17, 60–73 (2015).

  8. 8.

    Castillo, S. D. & Sanchez-Cespedes, M. The SOX family of genes in cancer development: biological relevance and opportunities for therapy. Expert Opin. Ther. Targets 16, 903–919 (2012).

  9. 9.

    Bar, Oz. M., Kumar, A., Elayyan, J., Reich, E., Binyamin, M., Kandel, L. et al. Acetylation reduces SOX9 nuclear entry and ACAN gene transactivation in human chondrocytes. Aging Cell 15, 499–508 (2016).

  10. 10.

    Liu, C., Liu, L., Chen, X., Cheng, J., Zhang, H., Shen, J. et al. Sox9 regulates self-renewal and tumorigenicity by promoting symmetrical cell division of cancer stem cells in hepatocellular carcinoma. Hepatology 64, 117–129 (2016).

  11. 11.

    Chakravarty, G., Moroz, K., Makridakis, N. M., Lloyd, S. A., Galvez, S. E., Canavello, P. R. et al. Prognostic significance of cytoplasmic SOX9 in invasive ductal carcinoma and metastatic breast cancer. Exp. Biol. Med. 236, 145–155 (2011).

  12. 12.

    Chakravarty, G., Rider, B. & Mondal, D. Cytoplasmic compartmentalization of SOX9 abrogates the growth arrest response of breast cancer cells that can be rescued by trichostatin A treatment. Cancer Biol. Ther. 11, 71–83 (2011).

  13. 13.

    Amano, K., Hata, K., Sugita, A., Takigawa, Y., Ono, K., Wakabayashi, M. et al. Sox9 family members negatively regulate maturation and calcification of chondrocytes through up-regulation of parathyroid hormone-related protein. Mol Biol Cell. 20, 4541–4551 (2009).

  14. 14.

    Dvir-Ginzberg, M., Gagarina, V., Lee, E. J. & Hall, D. J. Regulation of cartilage-specific gene expression in human chondrocytes by SirT1 and nicotinamide phosphoribosyltransferase. J Biol Chem. 283, 36300–36310 (2008)

  15. 15.

    Lefebvre. V. & Dvir-Ginzberg. M. SOX9 and the many facets of its regulation in the chondrocyte lineage. Connect Tissue Res. 58, 2–14 (2017).

  16. 16.

    Johnstone, R. W. Histone-deacetylase inhibitors: novel drugs for the treatment of cancer. Nat. Rev. Drug Discov. 1, 287–299 (2002).

  17. 17.

    Yang, X. J. & Seto, E. The Rpd3/Hda1 family of lysine deacetylases: from bacteria and yeast to mice and men. Nat. Rev. Mol. cell Biol. 9, 206–218 (2008).

  18. 18.

    Urbich, C., Rossig, L., Kaluza, D., Potente, M., Boeckel, J. N., Knau, A. et al. HDAC5 is a repressor of angiogenesis and determines the angiogenic gene expression pattern of endothelial cells. Blood 113, 5669–5679 (2009).

  19. 19.

    Duong, V., Bret, C., Altucci, L., Mai, A., Duraffourd, C., Loubersac, J. et al. Specific activity of class II histone deacetylases in human breast cancer cells. Mol. Cancer Res.: MCR 6, 1908–1919 (2008).

  20. 20.

    Huang, W. T., Tsai, Y. H., Chen, S. H., Kuo, C. W., Kuo, Y. L., Lee, K. T. et al. HDAC2 and HDAC5 up-regulations modulate survivin and mir-125a-5p expressions and promote hormone therapy resistance in estrogen receptor positive breast cancer cells. Front. Pharmacol. 8, 902 (2017).

  21. 21.

    Huang, Y., Vasilatos, S. N., Boric, L., Shaw, P. G., Davidson, N. E. Inhibitors of histone demethylation and histone deacetylation cooperate in regulating gene expression and inhibiting growth in human breast cancer cells. Breast Cancer Res. Treat. 131, 777–789 (2012).

  22. 22.

    Raha, P., Thomas, S., Thurn, K. T., Park, J., Munster, P. N. Combined histone deacetylase inhibition and tamoxifen induces apoptosis in tamoxifen-resistant breast cancer models, by reversing Bcl-2 overexpression. Breast Cancer Res. 17, 26 (2015).

  23. 23.

    Lu, M., Ding, K., Zhang, G., Yin, M., Yao, G., Tian, H. et al. MicroRNA-320a sensitizes tamoxifen-resistant breast cancer cells to tamoxifen by targeting ARPP-19 and ERRgamma. Sci. Rep. 5, 8735 (2015).

  24. 24.

    Shan, L., Zhou, X., Liu, X., Wang, Y., Su, D., Hou, Y. et al. FOXK2 Elicits Massive Transcription Repression and Suppresses the Hypoxic Response and Breast Cancer Carcinogenesis. Cancer cell 30, 708–722 (2016).

  25. 25.

    Gyorffy, B., Lanczky, A., Eklund, A. C., Denkert, C., Budczies, J., Li, Q. 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).

  26. 26.

    Nikolova, G. & Vilain, E. Mechanisms of disease: Transcription factors in sex determination-relevance to human disorders of sex development. Nat. Clin. Pract. Endocrinol. Metab. 2, 231–238 (2006).

  27. 27.

    Cao, C., Vasilatos, S. N., Bhargava, R., Fine, J. L., Oesterreich, S., Davidson, N. E. et al. Functional interaction of histone deacetylase 5 (HDAC5) and lysine-specific demethylase 1 (LSD1) promotes breast cancer progression. Oncogene 36, 133–145 (2017).

  28. 28.

    Jin, K., Park, S., Teo, W. W., Korangath, P., Cho, S. S., Yoshida, T. et al. HOXB7 Is an ERalpha Cofactor in the Activation of HER2 and Multiple ER Target Genes Leading to Endocrine Resistance. Cancer Discov. 5, 944–959 (2015).

  29. 29.

    Wan, J., Zhan, J., Li, S., Ma, J., Xu, W., Liu, C. et al. PCAF-primed EZH2 acetylation regulates its stability and promotes lung adenocarcinoma progression. Nucleic Acids Res. 43, 3591–3604 (2015).

  30. 30.

    Yuan, Z. L., Guan, Y. J., Chatterjee, D. & Chin, Y. E. Stat3 dimerization regulated by reversible acetylation of a single lysine residue. Science 307, 269–273 (2005).

  31. 31.

    Kruse, J. P. & Gu, W. SnapShot: p53 posttranslational modifications. Cell 133, 930–930 e931 (2008).

  32. 32.

    Wan, J., Xu, W., Zhan, J., Ma, J., Li, X., Xie, Y. et al. PCAF-mediated acetylation of transcriptional factor HOXB9 suppresses lung adenocarcinoma progression by targeting oncogenic protein JMJD6. Nucleic Acids Res. 44, 10662–10675 (2016).

  33. 33.

    Latham, J. A. & Dent, S. Y. Cross-regulation of histone modifications. Nat. Struct. Mol. Biol. 14, 1017–1024 (2007).

  34. 34.

    Vasilatos, S. N., Katz, T. A., Oesterreich, S., Wan, Y., Davidson, N. E. & Huang, Y. Crosstalk between lysine-specific demethylase 1 (LSD1) and histone deacetylases mediates antineoplastic efficacy of HDAC inhibitors in human breast cancer cells. Carcinogenesis 34, 1196–1207 (2013).

  35. 35.

    Chen, S., Yin, C., Lao, T., Liang, D., He, D., Wang, C. et al. AMPK-HDAC5 pathway facilitates nuclear accumulation of HIF-1alpha and functional activation of HIF-1 by deacetylating Hsp70 in the cytosol. Cell Cycle 14, 2520–2536 (2015).

  36. 36.

    Dean, M., Fojo, T. & Bates, S. Tumour stem cells and drug resistance. Nat. Rev. Cancer 5, 275–284 (2005).

  37. 37.

    Blumenschein, G. R. Jr., Kies, M. S., Papadimitrakopoulou, V. A., Lu, C., Kumar, A. J., Ricker, J. L. et al. Phase II trial of the histone deacetylase inhibitor vorinostat (Zolinza, suberoylanilide hydroxamic acid, SAHA) in patients with recurrent and/or metastatic head and neck cancer. Invest. New Drugs 26, 81–87 (2008).

  38. 38.

    Pili, R., Liu, G., Chintala, S., Verheul, H., Rehman, S., Attwood, K. et al. Combination of the histone deacetylase inhibitor vorinostat with bevacizumab in patients with clear-cell renal cell carcinoma: a multicentre, single-arm phase I/II clinical trial. Br. J. Cancer 116, 874–883 (2017).

  39. 39.

    Hynes, N. E. & Stoelzle, T. Key signalling nodes in mammary gland development and cancer: Myc. Breast Cancer Res. 11, 210 (2009).

Download references

Author information

Q.W., Y.X. and L.L. designed the research. Y.X., J.Z., W.L., W.Y., Q.L., Z.L. and H.Q. performed experiments, acquisition and analysis of data. Y.X., L.L. and Q.W. wrote the paper. J.G., X.G. and W.L. analysed the dataset of METABRIC. All authors have read and approved the paper.

Correspondence to Lihua Lai or Qingqing Wang.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethics approval and consent to participate

The human data involved in current study are obtained and analysed from the publicly archived datasets. There are no animal data in current study.

Funding

This work was supported by grants from the National Program on Key Basic Research Project (2014CB542101), National Natural Science Foundation of China (81230074, 81373115), Zhejiang Public Welfare Technology Research Project (GF19H160090) and Natural Science Foundation of Zhejiang Province (LY15H160012).

Consent to publish

Consent is obtained from all authors. All subjects gave written informed consent for publication.

Data availability

The datasets used and analysed during current study are available from the corresponding authors on reasonable request. The hyperlinks of publicly archived datasets involved in this study are as follows: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE9195; https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE9574; http://molonc.bccrc.ca/aparicio-lab/research/metabric/

Additional information

Note: This work is published under the standard license to publish agreement. After 12 months the work will become freely available and the license terms will switch to a Creative Commons Attribution 4.0 International (CC BY 4.0).

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Xue, Y., Lian, W., Zhi, J. et al. HDAC5-mediated deacetylation and nuclear localisation of SOX9 is critical for tamoxifen resistance in breast cancer. Br J Cancer (2019) doi:10.1038/s41416-019-0625-0

Download citation