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:

EZH2 suppresses IR-induced ferroptosis by forming a co-repressor complex with HIF-1α to inhibit ACSL4: Targeting EZH2 enhances radiosensitivity in KDM6A-deficient esophageal squamous carcinoma

Cell Death & Differentiation volume 32, pages 1026–1040 (2025)Cite this article

Subjects

Abstract

The mutation status of the lysine demethylase 6 A (KDM6A), a gene antagonist to Enhancer of zeste homolog 2 (EZH2), is closely related to the therapeutic efficacy of EZH2 inhibitors in several malignancies. However, the mutational landscape of KDM6A and the therapeutic targetability of EZH2 inhibitors in esophageal squamous carcinoma (ESCC) remain unreported. Here, we found that approximately 9.18% (9/98) of our study ESCC tissues had KDM6A mutations of which 7 cases resulted in a complete loss of expression and consequent loss of demethylase function. We found that KDM6A-deficient ESCC cells exhibited increased sensitivity to EZH2 inhibitor, and the radiosensitizing activity of EZH2 inhibitor was evident in KDM6A-dficient ESCC cells. Further transcriptome analysis revealed that ferroptosis is implicated in the radiosensitizing effect exerted by EZH2 inhibition on KDM6A-deficient ESCC cells. The following Chromatin Immunoprecipitation (ChIP), co-immunoprecipitation, and luciferase reporter assays demonstrated that in KDM6A-deficient ESCC cells, (1) Acyl-CoA Synthetase Long Chain Family Member 4 (ACSL4) is the target gene for EZH2 to regulate ferroptosis; (2) The IR-induced hypoxia inducible factor 1 subunit alpha (HIF-1α) is a predominant mediator of EZH2 to repress ACSL4; (3) the HRE7-8 regions of the ACSL4 promoter are required for the repressive function of EZH2 on ACSL4; (4) EZH2 regulates ACSL4 by forming a co-repressive complex with HIF-1α. Our study provides preclinical evidence supporting that EZH2 inhibitors may confer therapeutic benefit in KDM6A-deficient ESCC patients.

Access through your institution
Buy or subscribe

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

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

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

Fig. 1: A non-apoptotic type of cell death was involved in the regulation of EZH2 on radiosensitivity in KDM6A-deficient ESCC cells.
Fig. 2: EZH2 inhibition contributes to increase lipid peroxidation and MDA levels of KDM6A-deficient ESCC cells.
Fig. 3: EZH2 inhibition facilitates the morphological changes of mitochondrial damage and increases the expression levels of PTGS2.
Fig. 4: EZH2 inhibits IR-induced ferroptosis by downregulating ACSL4 in the guidance of HIF-1α.
Fig. 5: The repressive activity of EZH2 on ASCL4 is dependent on the presence of HRE7 and/or HRE8.
Fig. 6: Downregulation of EZH2 enhances the killing effect of IR on KDM6A-deficient ESCC cell xenografts.
Fig. 7: Proposed schematic representation of the molecular mechanism by which EZH2 modulates radiosensitivity in ESCC cells.

Similar content being viewed by others

Data availability

RNA-Seq data generated in this study is publicly available in the Gene Expression Omnibus (GEO) at GSE264079. Derived data supporting the findings of this study are available from the corresponding author upon request.

References

  1. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68:394–424.

    Article  PubMed  Google Scholar 

  2. Wu H, Yan Z, Zhang J, Sun X, Long H, Li D, et al. Correlation of CT texture changes with treatment response during radiation therapy for esophageal cancer. Int J Rad Oncol Biol Phys. 2019;105:E202–E3.

    Article  Google Scholar 

  3. Cohen DJ, Leichman L. Controversies in the treatment of local and locally advanced gastric and esophageal cancers. J Clin Oncol. 2015;33:1754–9.

    Article  PubMed  Google Scholar 

  4. Deng F, Zhou K, Cui W, Liu D, Ma Y. Clinicopathological significance of wnt/β-catenin signaling pathway in esophageal squamous cell carcinoma. Int J Clin Exper Pathol. 2015;8:3045.

    Google Scholar 

  5. Dixon SJ, Lemberg KM, Lamprecht MR, Skouta R, Zaitsev EM, Gleason CE, et al. Ferroptosis: an iron-dependent form of nonapoptotic cell death. Cell. 2012;149:1060–72.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Stockwell BR, Friedmann Angeli JP, Bayir H, Bush AI, Conrad M, Dixon SJ, et al. Ferroptosis: a regulated cell death nexus linking metabolism, redox biology, and disease. Cell. 2017;171:273–85.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Doll S, Proneth B, Tyurina YY, Panzilius E, Kobayashi S, Ingold I, et al. ACSL4 dictates ferroptosis sensitivity by shaping cellular lipid composition. Nat Chem Biol. 2017;13:91–8.

    Article  CAS  PubMed  Google Scholar 

  8. Yuan Z-h, Liu T, Wang H, Xue L-x, Wang J-j. Fatty acids metabolism: the bridge between ferroptosis and ionizing radiation. Front Cell Dev Biol. 2021;9:675617.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Koppula P, Zhuang L, Gan B. Cystine transporter SLC7A11/xCT in cancer: ferroptosis, nutrient dependency, and cancer therapy. Protein Cell. 2021;12:599–620.

    Article  CAS  PubMed  Google Scholar 

  10. Ye LF, Chaudhary KR, Zandkarimi F, Harken AD, Kinslow CJ, Upadhyayula PS, et al. Radiation-induced lipid peroxidation triggers ferroptosis and synergizes with ferroptosis inducers. ACS Chem Biol. 2020;15:469–84.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Kim EJ, Lee Y-J, Kang S, Lim Y-B. Ionizing radiation activates PERK/eIF2α/ATF4 signaling via ER stress-independent pathway in human vascular endothelial cells. Int J Radiat Biol. 2014;90:306–12.

    Article  CAS  PubMed  Google Scholar 

  12. Lang X, Green MD, Wang W, Yu J, Choi JE, Jiang L, et al. Radiotherapy and immunotherapy promote tumoral lipid oxidation and ferroptosis via synergistic repression of SLC7A11. Cancer Discov. 2019;9:1673–85.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Lei G, Zhang Y, Koppula P, Liu X, Zhang J, Lin SH, et al. The role of ferroptosis in ionizing radiation-induced cell death and tumor suppression. Cell Res. 2020;30:146–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Duan R, Du W, Guo W. EZH2: a novel target for cancer treatment. J Hematol Oncol. 2020;13:104.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Liu N, Konuma T, Sharma R, Wang D, Zhao N, Cao L, et al. Histone H3 lysine 27 crotonylation mediates gene transcriptional repression in chromatin. Mol Cell. 2023;83:2206–21.e11.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. He LR, Liu MZ, Li BK, Jia WH, Zhang Y, Liao YJ, et al. High expression of EZH2 is associated with tumor aggressiveness and poor prognosis in patients with esophageal squamous cell carcinoma treated with definitive chemoradiotherapy. Int J Cancer. 2010;127:138–47.

    Article  CAS  PubMed  Google Scholar 

  17. Huang X, Yan J, Zhang M, Wang Y, Chen Y, Fu X, et al. Targeting epigenetic crosstalk as a therapeutic strategy for EZH2-aberrant solid tumors. Cell. 2018;175:186–99.e19.

    Article  CAS  PubMed  Google Scholar 

  18. Li C, Wang Y, Gong Y, Zhang T, Huang J, Tan Z, et al. Finding an easy way to harmonize: a review of advances in clinical research and combination strategies of EZH2 inhibitors. Clin Epigenetics. 2021;13:62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Pugh TJ, Weeraratne SD, Archer TC, Pomeranz Krummel DA, Auclair D, Bochicchio J, et al. Medulloblastoma exome sequencing uncovers subtype-specific somatic mutations. Nature. 2012;488:106–10.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. van Haaften G, Dalgliesh GL, Davies H, Chen L, Bignell G, Greenman C, et al. Somatic mutations of the histone H3K27 demethylase gene UTX in human cancer. Nat Genet. 2009;41:521–3.

    Article  PubMed  PubMed Central  Google Scholar 

  21. Jankowska AM, Makishima H, Tiu RV, Szpurka H, Huang Y, Traina F, et al. Mutational spectrum analysis of chronic myelomonocytic leukemia includes genes associated with epigenetic regulation: UTX, EZH2, and DNMT3A. Blood. 2011;118:3932–41.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Ler LD, Ghosh S, Chai X, Thike AA, Heng HL, Siew EY, et al. Loss of tumor suppressor KDM6A amplifies PRC2-regulated transcriptional repression in bladder cancer and can be targeted through inhibition of EZH2. Sci Transl Med. 2017;9:984–5.

    Article  Google Scholar 

  23. Ezponda T, Dupere-Richer D, Will CM, Small EC, Varghese N, Patel T, et al. UTX/KDM6A loss enhances the malignant phenotype of multiple myeloma and sensitizes cells to EZH2 inhibition. Cell Rep. 2017;21:628–40.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Gao YB, Chen ZL, Li JG, Hu XD, Shi XJ, Sun ZM, et al. Genetic landscape of esophageal squamous cell carcinoma. Nat Genet. 2014;46:1097–102.

    Article  CAS  PubMed  Google Scholar 

  25. Lin DC, Hao JJ, Nagata Y, Xu L, Shang L, Meng X, et al. Genomic and molecular characterization of esophageal squamous cell carcinoma. Nat Genet. 2014;46:467–73.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Cancer Genome Atlas Research N, Analysis Working Group: Asan U, Agency BCC, Brigham, Women’s H, Broad I. et al. Integrated genomic characterization of oesophageal carcinoma. Nature. 2017;541:169–75.

    Article  Google Scholar 

  27. Tong ZT, Cai MY, Wang XG, Kong LL, Mai SJ, Liu YH, et al. EZH2 supports nasopharyngeal carcinoma cell aggressiveness by forming a co-repressor complex with HDAC1/HDAC2 and Snail to inhibit E-cadherin. Oncogene. 2012;31:583–94.

    Article  CAS  PubMed  Google Scholar 

  28. Cai MY, Tong ZT, Zheng F, Liao YJ, Wang Y, Rao HL, et al. EZH2 protein: a promising immunomarker for the detection of hepatocellular carcinomas in liver needle biopsies. Gut. 2011;60:967–76.

    Article  CAS  PubMed  Google Scholar 

  29. Tong Z, Fang W, Xu M, Xia Y, Wang R, Li Y, et al. DAB2IP predicts treatment response and prognosis of ESCC patients and modulates its radiosensitivity through enhancing IR-induced activation of the ASK1-JNK pathway. Cancer Cell Int. 2022;22:106.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Sak A, Stuschke M. Use of gammaH2AX and other biomarkers of double-strand breaks during radiotherapy. Semin Radiat Oncol. 2010;20:223–31.

    Article  PubMed  Google Scholar 

  31. Schultz LB, Chehab NH, Malikzay A, Halazonetis TD. p53 binding protein 1 (53BP1) is an early participant in the cellular response to DNA double-strand breaks. J Cell Biol. 2000;151:1381–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Burma S, Chen BP, Murphy M, Kurimasa A, Chen DJ. ATM phosphorylates histone H2AX in response to DNA double-strand breaks. J Biol Chem. 2001;276:42462–7.

    Article  CAS  PubMed  Google Scholar 

  33. Meyn RE, Milas L, Ang KK. The role of apoptosis in radiation oncology. Int J Radiat Biol. 2009;85:107–15.

    Article  CAS  PubMed  Google Scholar 

  34. Muller J, Kassis JA. Polycomb response elements and targeting of Polycomb group proteins in Drosophila. Curr Opin Genet Dev. 2006;16:476–84.

    Article  PubMed  Google Scholar 

  35. Wilkinson F, Pratt H, Atchison ML. PcG recruitment by the YY1 REPO domain can be mediated by Yaf2. J Cellular Biochem. 2010;109:478–86.

    Article  CAS  Google Scholar 

  36. Jiang Y Fasting alters histone methylation in paraventricular nucleus of chick through regulating of polycomb repressive complex 2: Virginia Tech; 2013.

  37. Quan J, Bode AM, Luo X. ACSL family: The regulatory mechanisms and therapeutic implications in cancer. Eur J Pharmacol. 2021;909:174397.

    Article  CAS  PubMed  Google Scholar 

  38. Moeller BJ, Dewhirst MW. HIF-1 and tumour radiosensitivity. Br J Cancer. 2006;95:1–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Masoud GN, Li W. HIF-1alpha pathway: role, regulation and intervention for cancer therapy. Acta Pharm Sin B. 2015;5:378–89.

    Article  PubMed  PubMed Central  Google Scholar 

  40. Li Y, Zhao X, Tang H, Zhong Z, Zhang L, Xu R, et al. Effects of YC-1 on hypoxia-inducible factor 1 alpha in hypoxic human bladder transitional carcinoma cell line T24 cells. Urol Int. 2012;88:95–101.

    Article  CAS  PubMed  Google Scholar 

  41. Ouyang C, Zhang J, Lei X, Xie Z, Liu X, Li Y, et al. Advances in antitumor research of HIF-1α inhibitor YC-1 and its derivatives. Bioog Chem. 2023;133:106400.

  42. Masoud GN, Li W. HIF-1α pathway: role, regulation and intervention for cancer therapy. Acta Pharm Sin B. 2015;5:378–89.

    Article  PubMed  PubMed Central  Google Scholar 

  43. Wu Q, You L, Nepovimova E, Heger Z, Wu W, Kuca K, et al. Hypoxia-inducible factors: master regulators of hypoxic tumor immune escape. J Hematol Oncol. 2022;15:77.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Liu GZ, Xu XW, Tao SH, Gao MJ, Hou ZH. HBx facilitates ferroptosis in acute liver failure via EZH2 mediated SLC7A11 suppression. J Biomed Sci. 2021;28:67.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Yi C, Li G, Wang W, Sun Y, Zhang Y, Zhong C, et al. Disruption of YY1-EZH2 interaction using synthetic peptides inhibits breast cancer development. Cancers. 2021;13:2402.

  46. Moeller BJ, Cao Y, Li CY, Dewhirst MW. Radiation activates HIF-1 to regulate vascular radiosensitivity in tumors: role of reoxygenation, free radicals, and stress granules. Cancer Cell. 2004;5:429–41.

    Article  CAS  PubMed  Google Scholar 

  47. Lee TI, Jenner RG, Boyer LA, Guenther MG, Levine SS, Kumar RM, et al. Control of developmental regulators by Polycomb in human embryonic stem cells. Cell. 2006;125:301–13.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Boyer LA, Plath K, Zeitlinger J, Brambrink T, Medeiros LA, Lee TI, et al. Polycomb complexes repress developmental regulators in murine embryonic stem cells. Nature. 2006;441:349–53.

    Article  CAS  PubMed  Google Scholar 

  49. Kim SH, Joshi K, Ezhilarasan R, Myers TR, Siu J, Gu C, et al. EZH2 protects glioma stem cells from radiation-induced cell death in a MELK/FOXM1-dependent manner. Stem Cell Rep. 2015;4:226–38.

    Article  CAS  Google Scholar 

  50. Zhang X, Ma X, Wang Q, Kong Z. EZH2 targeting to improve the sensitivity of acquired radio-resistance bladder cancer cells. Transl Oncol. 2022;16:101316.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We are grateful to Dr. Jiabing Fan (University of California, Los Angeles) for his invaluable comments and extensive manuscript editing. We also thank LetPub (www.letpub.com) for its linguistic assistance during the preparation of this manuscript.

Funding

This study was supported by the University Natural Science Research Project of Anhui Province (Grant Number: KJ2021A0295), the National Natural Science Foundation of China (Grant Number: 81201743), the Health Research Program of Anhui (Grant Number: AHWJ2023BAc20040), and the Natural Science Foundation of Anhui Province, China (Grant Number: 1908085QA27).

Author information

Author notes

  1. Yeye Xia

    Present address: Department of Oncology, Chengdu Fifth People’s Hospital, Chengdu, China

  2. Qianqian Zhu

    Present address: Department of Radiation Oncology, Fuyang Tumour Hospital, Fuyang, China

  3. These authors contributed equally: Guizhen Pan, Yeye Xia, Mengyu Hao.

Authors and Affiliations

  1. Department of Radiation Oncology, the First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China

    Guizhen Pan, Yeye Xia, Mengyu Hao, Jiahao Guan, Qianqian Zhu, Tianqi Zha, Zhenfeng Zhao, Xiangcun Chen & Zhuting Tong

  2. Department of Radiation Oncology, the Chest Hospital of Anhui Province, Hefei, Anhui, China

    Lei Sheng

  3. Anhui Public Health Clinical Center, Hefei, Anhui, China

    Zhenfeng Zhao

  4. Department of Thoracic Surgery, the First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China

    Huaguang Pan

  5. Department of Electrocardiography, the First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China

    Weiyang Fang

  6. Department of Gastroenterology, the First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China

    Xiaoyong Xu

  7. The Fifth Clinical College of Anhui Medical University, Hefei, Anhui, China

    Shuguang Zhou

Authors
  1. Guizhen Pan
    View author publications

    Search author on:PubMed Google Scholar

  2. Yeye Xia
    View author publications

    Search author on:PubMed Google Scholar

  3. Mengyu Hao
    View author publications

    Search author on:PubMed Google Scholar

  4. Jiahao Guan
    View author publications

    Search author on:PubMed Google Scholar

  5. Qianqian Zhu
    View author publications

    Search author on:PubMed Google Scholar

  6. Tianqi Zha
    View author publications

    Search author on:PubMed Google Scholar

  7. Lei Sheng
    View author publications

    Search author on:PubMed Google Scholar

  8. Zhenfeng Zhao
    View author publications

    Search author on:PubMed Google Scholar

  9. Huaguang Pan
    View author publications

    Search author on:PubMed Google Scholar

  10. Weiyang Fang
    View author publications

    Search author on:PubMed Google Scholar

  11. Xiaoyong Xu
    View author publications

    Search author on:PubMed Google Scholar

  12. Xiangcun Chen
    View author publications

    Search author on:PubMed Google Scholar

  13. Shuguang Zhou
    View author publications

    Search author on:PubMed Google Scholar

  14. Zhuting Tong
    View author publications

    Search author on:PubMed Google Scholar

Contributions

ZT conceived and designed the study. GP, YX, and MH performed most of the experiments. WF, JG, LS, SZ, and TZ assisted in the performance of the experiments. HP, XX, QZ, and XC collected patient samples. ZT, GP, YX, MH, and JG contributed to data analysis. ZT, GP, and YX prepared and revised the figures, and drafted the manuscript with assistance from JG and MH. ZT, XC, and QZ provided funding. ZT supervised the research. All authors read and approved the final version of the manuscript.

Corresponding author

Correspondence to Zhuting Tong.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethical approval and consent to participate

Approval for human tissue samples utilization was granted by the Biomedical Ethics Committee of Anhui Medical University (5101745), following the guidelines of Declaration of Helsinki. All patients authorized the use of their specimens by written informed consent. All animal experiments were carried out with the approval of the Experimental Animal Ethical Committee of Anhui Medical University (LLSC20210863) and performed according to institutional guidelines.

Additional information

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

Supplementary information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Pan, G., Xia, Y., Hao, M. et al. EZH2 suppresses IR-induced ferroptosis by forming a co-repressor complex with HIF-1α to inhibit ACSL4: Targeting EZH2 enhances radiosensitivity in KDM6A-deficient esophageal squamous carcinoma. Cell Death Differ 32, 1026–1040 (2025). https://doi.org/10.1038/s41418-025-01451-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue date:

  • DOI: https://doi.org/10.1038/s41418-025-01451-5

Search

Advanced search

Quick links