EZH2 regulates expression of FOXC1 by mediating H3K27me3 in breast cancers

Abstract

Triple-negative breast cancer (TNBC) is characterized by low expression of human epidermal growth factor receptor-2 (HER2), estrogen receptor (ER), and progesterone receptor (PR), which is the most aggressive subtype with poor outcome among breast cancers. The underlying mechanisms of TNBC remain unclear and there is a lack of biomarkers. In this study we conducted an in silico assay and found that FOXC1 was highly expressed in ER/PR/HER2 breast cancers, which was confirmed by qRT-PCR, immunohistochemistry, and Western blot analysis. FOXC1 was more highly expressed in TNBCs than the other breast cancers. Kaplan–Meier plotter revealed that expression of FOXC1 was associated with overall survival (OS) of patients with breast cancers. Expression of FOXC1 was reversely associated with level of H3K27me3, which was methylated by EZH2. In MCF-7 and T47D cells, inhibition of EZH2 by DZNeP or GSK343 concentration- and time-dependently increased expression of FOXC1. Finally, we demonstrated that the expression of FOXC1 was associated with resistance of doxorubicin treatment of breast cancer cells. In conclusion, these results suggest that FOXC1 may be a potential biomarker or drug target for TNBCs, and that downregulation of FOXC1 could have therapeutic value in treatment of TNBCs.

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Fig. 1: FOXC1 mRNA was highly expressed in TNBCs.
Fig. 2: FOXC1 protein was highly expressed in TNBCs and associated with breast cancer progression.
Fig. 3: Expression of FOXC1 was associated with a poor prognosis in breast cancer patients.
Fig. 4: Expression of FOXC1 was reversibly related to histone methylation (H3K27me3) by EZH2.
Fig. 5: Overexpression of EZH2 inhibited the expression of FOXC1 by increasing H3K27me3 level.
Fig. 6: Inhibition of EZH2 in MCF-7 cells induced the expression of FOXC1.
Fig. 7: Inhibition of demethylases KDM6A and KDM6B did not alter the expression of FOXC1 in breast cancer cells.
Fig. 8: FOXC1 overexpression was associated with doxorubicin (Dox) resistance.
Fig. 9: Proposed mechanism: EZH2 regulates the expression of FOXC1 by mediating H3K27me3 in breast cancers.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 81573454 to JHW, No. 81703536 to WL) and the Beijing Natural Science Foundation of Beijing Municipality (7172142). This work was also supported by the CAMS Innovation Fund for Medical Sciences (2016-I2M-3-007) and the Technology Major Projects for “Major New Drugs Innovation and Development” (2018ZX09711001-005-025, 2018ZX09711001-012).

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JHW and GHD developed the hypothesis, designed the experiments, and revised the manuscript. XJZ and WL conducted all experiments and wrote the main manuscript. JYL, LWR, SWL, and XMZ collected data and performed the statistical analysis. JY provided the study materials and patients. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Jin-hua Wang or Guan-hua Du.

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The authors declare no competing interests.

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Zheng, X., Li, W., Yi, J. et al. EZH2 regulates expression of FOXC1 by mediating H3K27me3 in breast cancers. Acta Pharmacol Sin (2020). https://doi.org/10.1038/s41401-020-00543-x

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Keywords

  • TNBC
  • FOXC1
  • EZH2
  • H3K27me3
  • drug resistance

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