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Cellular and Molecular Biology

HIF-1α stimulates the progression of oesophageal squamous cell carcinoma by activating the Wnt/β-catenin signalling pathway



This study aimed to clarify the significance of the crosstalk between hypoxia-inducible factor-1α (HIF-1α) and the Wnt/β-catenin pathway in oesophageal squamous cell carcinoma (ESCC).


The oncogenic role of HIF-1α in ESCC was investigated using in vitro and in vivo assays. The clinicopathological significance of HIF-1α, β-catenin and TCF4/TCF7L2 in ESCC were evaluated using quantitative real-time PCR and immunohistochemistry.


The expression level of HIF-1α, β-catenin, and TCF4/TCF7L2 in T.Tn and TE1 cell lines were elevated under hypoxia in vitro. HIF-1α knockdown suppressed proliferation, migration/invasion and epithelial–mesenchymal transition (EMT) progression, induced G0/G1 cell cycle arrest, promoted apoptosis and inhibited 5-fluorouracil chemoresistance in vitro. In vivo assays showed that HIF-1α is essential in maintaining tumour growth, angiogenesis, and 5-fluorouracil chemoresistance. Mechanically, we identified the complex between HIF-1α and β-catenin, HIF-1α can directly bind to the promoter region of TCF4/TCF7L2. The mRNA level of HIF-1α, β-catenin and TCF4/TCF7L2 were increased in ESCC tumour tissues compared to the corresponding non-tumour tissues. High levels of HIF-1α and TCF4/TCF7L2 expression were correlated with aggressive phenotypes and poor prognosis in ESCC patients.


HIF-1α serves as an oncogenic transcriptional factor in ESCC, probably by directly targeting TCF4/TCF7L2 and activating the Wnt/β-catenin pathway.

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Fig. 1: 1% O2 or CoCl2 induced hypoxia and HIF-1α shRNA regulating HIF-1α expression in ESCC cell lines.
Fig. 2: The role of HIF-1α in ESCC cell proliferation, migration, invasion, apoptosis and cell cycle in vitro.
Fig. 3: HIF-1α was involved in hypoxia-induced EMT activation in T.Tn and TE1 cell lines.
Fig. 4: HIF-1α mediated the resistance of ESCC cell lines to 5-FU in vitro and in vivo.
Fig. 5: Crosstalk between HIF-1α and Wnt/β-catenin signalling pathway in ESCC cell lines.
Fig. 6: HIF-1α expression was associated with TCF4/TCF7L2 expression in ESCC tissues, which was correlated with poor prognosis.

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Data availability

All data required to evaluate the findings in the paper is available in the paper and supplementary materials.


  1. Pennathur A, Gibson MK, Jobe BA, Luketich JD. Oesophageal carcinoma. Lancet. 2013;381:400–12.

    Article  Google Scholar 

  2. Lagergren J, Smyth E, Cunningham D, Lagergren P. Oesophageal cancer. Lancet. 2017;390:2383–96.

    Article  Google Scholar 

  3. Wilson WR, Hay MP. Targeting hypoxia in cancer therapy. Nat Rev Cancer. 2011;11:393–410.

    Article  CAS  Google Scholar 

  4. Ke Q, Costa M. Hypoxia-inducible factor-1 (HIF-1). Pharmacol. 2006;70:1469–80.

    CAS  Google Scholar 

  5. Hajizadeh F, Okoye I, Esmaily M, Ghasemi Chaleshtari M, Masjedi A, Azizi G, et al. Hypoxia inducible factors in the tumor microenvironment as therapeutic targets of cancer stem cells. Life Sci. 2019;

  6. Heddleston JM, Li Z, Lathia JD, Bao S, Hjelmeland AB, Rich JN. Hypoxia inducible factors in cancer stem cells. Br J Cancer. 2010;

  7. Wiesener MS, Jürgensen JS, Rosenberger C, Scholze CK, Hörstrup JH, Warnecke C, et al. Widespread hypoxia-inducible expression of HIF-2alpha in distinct cell populations of different organs. FASEB J. 2003.

  8. Keith B, Simon MC. Hypoxia-inducible factors, stem cells, and cancer. Cell. 2007;129:465–72.

    Article  CAS  Google Scholar 

  9. Gonzalez FJ, Xie C, Jiang C. The role of hypoxia-inducible factors in metabolic diseases. Nat Rev Endocrinol. 2018;15:21–32.

    Article  Google Scholar 

  10. Eubank TD, Roda JM, Liu H, O’Neil T, Marsh CB. Opposing roles for HIF-1α and HIF-2α in the regulation of angiogenesis by mononuclear phagocytes. Blood. 2011.

  11. Zhang Y, Wang X. Targeting the Wnt/β-catenin signaling pathway in cancer. J Hematol Oncol. 2020;13:165.

    Article  Google Scholar 

  12. Boso D, Rampazzo E, Zanon C, Bresolin S, Maule F, Porcù E, et al. HIF-1α/Wnt signaling-dependent control of gene transcription regulates neuronal differentiation of glioblastoma stem cells. Theranostics. 2019;9:4860–77.

    Article  CAS  Google Scholar 

  13. Jiang N, Zou C, Zhu Y, Luo Y, Chen L, et al. HIF-1ɑ-regulated miR-1275 maintains stem cell-like phenotypes and promotes the progression of LUAD by simultaneously activating Wnt/β-catenin and Notch signaling. Theranostics. 2020;10:2553–70.

    Article  CAS  Google Scholar 

  14. Scholten DJ 2nd, Timmer CM, Peacock JD, Pelle DW, Williams BO, Steensma MR. Down regulation of Wnt signaling mitigates hypoxia-induced chemoresistance in human osteosarcoma cells. PLoS ONE. 2014;9:e111431.

    Article  Google Scholar 

  15. Sashiyama H, Shino Y, Kawamata Y, Tomita Y, Ogawa N, Shimada H, et al. Immortalization of human esophageal keratinocytes by E6 and E7 of human papillomavirus type 16. Int J Oncol. 2001;19:97–103.

    CAS  PubMed  Google Scholar 

  16. Ji Z, Yang G, Shahzidi S, Tkacz-Stachowska K, Suo Z, Nesland JM. Induction of hypoxia-inducible factor-1alpha overexpression by cobalt chloride enhances cellular resistance to photodynamic therapy. Cancer Lett. 2006.

  17. Dai M, Cui P, Yu M, Han J, Li H, Xiu R. Melatonin modulates the expression of VEGF and HIF-1 alpha induced by CoCl2 in cultured cancer cells. J Pineal Res. 2008;44:121–6.

    Article  CAS  Google Scholar 

  18. Toyozumi T, Hoshino I, Takahashi M, Usui A, Akutsu Y, Hanari N, et al. Fra-1 Regulates the expression of HMGA1, which is associated with a poor prognosis in human esophageal squamous cell carcinoma. Ann Surg Oncol. 2017.

  19. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 2001;25:402–8.

    Article  CAS  Google Scholar 

  20. Liu DS, Read M, Cullinane C, Azar WJ, Fennell CM, Montgomery KG, et al. APR-246 potently inhibits tumour growth and overcomes chemoresistance in preclinical models of oesophageal adenocarcinoma. Gut. 2015;64:1506–16.

    Article  CAS  Google Scholar 

  21. Weidner N, Semple JP, Welch WR, Folkman J. Tumor angiogenesis and metastasis-correlation in invasive breast carcinoma. N Engl J Med. 1991;324:1–8.

    Article  CAS  Google Scholar 

  22. Pastushenko I, Blanpain C. EMT transition states during tumor progression and metastasis. Trends Cell Biol. 2019;

  23. Mazumdar J, O’Brien WT, Johnson RS, LaManna JC, Chavez JC, Klein PS, et al. O2 regulates stem cells through Wnt/β-catenin signalling. Nat Cell Biol. 2010;12:1007–13.

    Article  CAS  Google Scholar 

  24. Liu HL, Liu D, Ding GR, Liao PF, Zhang JW. Hypoxia-inducible factor-1α and Wnt/β-catenin signaling pathways promote the invasion of hypoxic gastric cancer cells. Mol Med Rep. 2015;

  25. Hong CF, Chen WY, Wu CW. Upregulation of Wnt signaling under hypoxia promotes lung cancer progression. Oncol Rep. 2017;

  26. Xu W, Zhou W, Cheng M, Wang J, Liu Z, He S, et al. Hypoxia activates Wnt/β-catenin signaling by regulating the expression of BCL9 in human hepatocellular carcinoma. Sci Rep. 2017.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Muz B, de la Puente P, Azab F, Azab AK. The role of hypoxia in cancer progression, angiogenesis, metastasis, and resistance to therapy. Hypoxia. 2015;3:83–92.

    Article  Google Scholar 

  28. Pouysségur J, Dayan F, Mazure NM. Hypoxia signalling in cancer and approaches to enforce tumour regression. Nature. 2006;441:437–43.

    Article  Google Scholar 

  29. Jing X, Yang F, Shao C, Wei K, Xie M, Shen H. Role of hypoxia in cancer therapy by regulating the tumor microenvironment. Mol Cancer. 2019;18:157.

    Article  Google Scholar 

  30. Belisario DC, Kopecka J, Pasino M, Akman M, De Smaele E, Donadelli M. Hypoxia dictates metabolic rewiring of tumors: implications for chemoresistance. Cells. 2020;9:2598.

    Article  CAS  Google Scholar 

  31. Kaidi A, Williams AC, Paraskeva C. Interaction between beta-catenin and HIF-1 promotes cellular adaptation to hypoxia. Nat Cell Biol. 2007;

  32. Rezaeian AH, Li CF, Wu CY, Zhang X, Delacerda J, You MJ, et al. A hypoxia-responsive TRAF6-ATM-H2AX signalling axis promotes HIF1α activation, tumorigenesis and metastasis. Nat Cell Biol. 2017;

  33. Liu M, Zhong J, Zeng Z, Huang K, Ye Z, Deng S, et al. Hypoxia-induced feedback of HIF-1α and lncRNA-CF129 contributes to pancreatic cancer progression through stabilization of p53 protein. Theranostics. 2019;9:4795–810.

    Article  CAS  Google Scholar 

  34. Alsaab HO, Sau S, Alzhrani RM, Cheriyan VT, Polin LA, Vaishampayan U. Tumor hypoxia directed multimodal nanotherapy for overcoming drug resistance in renal cell carcinoma and reprogramming macrophages. Biomaterials. 2018;

  35. Kitajima S, Lee KL, Hikasa H, Sun W, Huang RY, Yang H, et al. Hypoxia-inducible factor-1α promotes cell survival during ammonia stress response in ovarian cancer stem-like cells. Oncotarget. 2017;8:114481–94.

    Article  Google Scholar 

  36. Koukourakis MI, Giatromanolaki A, Skarlatos J, Corti L, Blandamura S, Piazza, et al. Hypoxia inducible factor (HIF-1a and HIF-2a) expression in early esophageal cancer and response to photodynamic therapy and radiotherapy. Cancer Res. 2001;61:1830–2.

  37. Ishiguro H, Wakasugi T, Terashita Y, Sakamoto N, Tanaka T, Sagawa H, et al. Nuclear expression of TCF4/TCF7L2 is correlated with poor prognosis in patients with esophageal squamous cell carcinoma. Cell Mol Biol Lett. 2016;21:5.

    Article  Google Scholar 

  38. Rana NK, Singh P, Koch B. CoCl2 simulated hypoxia induce cell proliferation and alter the expression pattern of hypoxia associated genes involved in angiogenesis and apoptosis. Biol Res. 2019;52:12.

    Article  Google Scholar 

  39. Vengellur A, Phillips JM, Hogenesch JB, LaPres JJ. Gene expression profiling of hypoxia signaling in human hepatocellular carcinoma cells. Physiol Genomics. 2005;

  40. Borcar A, Menze MA, Toner M, Hand SC. Metabolic preconditioning of mammalian cells: mimetic agents for hypoxia lack fidelity in promoting phosphorylation of pyruvate dehydrogenase. Cell Tissue Res. 2013;

  41. Heerboth S, Housman G, Leary M, Longacre M, Byler S, Lapinska K, et al. EMT and tumor metastasis. Clin Transl Med. 2015;4:6.

    Article  Google Scholar 

  42. Dou C, Zhou Z, Xu Q, Liu Z, Zeng Y, Wang Y, et al. Hypoxia-induced TUFT1 promotes the growth and metastasis of hepatocellular carcinoma by activating the Ca2+/PI3K/AKT pathway. Oncogene. 2019;

  43. Luan S, Zeng X, Zhang C, Qiu J, Yang Y, Mao C, et al. Advances in drug resistance of esophageal cancer: from the perspective of tumor microenvironment. Front Cell Dev Biol. 2021;9:664816.

    Article  Google Scholar 

  44. Wu ZQ, Li XY, Hu CY, Ford M, Kleer CG, Weiss SJ. Canonical Wnt signaling regulates Slug activity and links epithelial-mesenchymal transition with epigenetic breast cancer 1, early onset (BRCA1) repression. Proc Natl Acad Sci USA. 2012;

  45. Zhang Q, Bai X, Chen W, Ma T, Hu Q, Liang C, et al. Wnt/β-catenin signaling enhances hypoxia-induced epithelial-mesenchymal transition in hepatocellular carcinoma via crosstalk with hif-1α signaling. Carcinogenesis. 2013;

  46. He TC, Sparks AB, Rago C, Hermeking H, Zawel L, da Costa LT, et al. Identification of c-MYC as a target of the APC pathway. Science. 1998;281:1509–12.

    Article  CAS  Google Scholar 

  47. Ju X, Casimiro MC, Gormley M, Meng H, Jiao X, Katiyar S, et al. Identification of a cyclin D1 network in prostate cancer that antagonizes epithelial-mesenchymal restraint. Cancer Res. 2014;

  48. Nusse R, Clevers H. Wnt/β-catenin signaling, disease, and emerging therapeutic modalities. Cell. 2017;169:985–99.

    Article  CAS  Google Scholar 

  49. He G, Guan X, Chen X, Wang Y, Luo C, Zhang B. Expression and splice variant analysis of human TCF4 transcription factor in esophageal cancer. J Cancer. 2015;6:333–41.

    Article  Google Scholar 

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We sincerely thank the first author’s wife, Dr. Xiayun Wan, for her psychological counselling during the author’s writing period.


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Authors and Affiliations



KT, TT, KM, HS, MK, SE, YM, MU, KH and HM conceived this study. KT performed all the experiments. HS, MT, NS, RO, KK, SH and JH monitored the progress of the experiment, provided comments and helped write the manuscript. KT performed the statistics and wrote the manuscript. TT revised the manuscript. HM approved the final version. All authors have reviewed the manuscript and all have approved the submission.

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Correspondence to Takeshi Toyozumi.

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The authors declare no competing interests.

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This research was performed in accordance with the Declaration of Helsinki. Our animal research was approved by the Animal Care and Use Committee of Chiba University (No. A3-22) and follows the policies for institutional animal care. The clinical analyses in this study were authorised by the Institutional Review Board of Chiba University (No. 1120-942) and written informed consent was obtained from all participants.

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Tang, K., Toyozumi, T., Murakami, K. et al. HIF-1α stimulates the progression of oesophageal squamous cell carcinoma by activating the Wnt/β-catenin signalling pathway. Br J Cancer 127, 474–487 (2022).

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