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Direct regulation of TWIST by HIF-1α promotes metastasis

Abstract

Stabilization of the hypoxia-inducible factor-1α (HIF-1α) transcription complex, caused by intratumoural hypoxia, promotes tumour progression and metastasis, leading to treatment failure and mortality in different types of human cancers. The transcription factor TWIST is a master regulator of gastrulation and mesoderm-specification and was implicated recently as an essential mediator of cancer metastasis. Notably, HIF-1α- and TWIST-null mice show similarities in their phenotypes. Here, we have shown that hypoxia or overexpression of HIF-1α promotes epithelial–mesenchymal transition (EMT) and metastastic phenotypes. We also found that HIF-1 regulates the expression of TWIST by binding directly to the hypoxia-response element (HRE) in the TWIST proximal promoter. However, siRNA-mediated repression of TWIST in HIF-1α-overexpressing or hypoxic cells reversed EMT and metastastic phenotypes. Co-expression of HIF-1α, TWIST and Snail in primary tumours of patients with head and neck cancers correlated with metastasis and the worst prognosis. These results provide evidence of a key signalling pathway involving HIF-1α and TWIST that promotes metastasis in response to intratumoural hypoxia.

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Figure 1: Hypoxia or overexpression of HIF-1α(ΔODD) induces EMT and causes increases in migration, invasion and tumour size.
Figure 2: Constitutive expression of HIF-1α(ΔODD) in FaDu cells promotes metastasis and local invasion in vivo.
Figure 3: Hypoxia or constitutive expression of HIF-1α(ΔODD) upregulates TWIST expression.
Figure 4: HIF-1 upregulates TWIST expression by direct binding to the HRE in the proximal promoter of TWIST gene.
Figure 5: Knockdown of TWIST in FaDu–HIF1α(ΔODD) or H1299 cells reverses EMT and metastasis.
Figure 6: Restoration of TWIST expression or repression of Snail in respective cell lines.
Figure 7: Co-expression of CA IX, HIF-1α, TWIST and Snail in a HNSCC case.
Figure 8: Co-expression of HIF-1α, TWIST and Snail correlates with metastasis of HNSCC.

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References

  1. Gupta, G. P. & Massague, J. Cancer metastasis: building a framework. Cell 127, 679–695 (2006).

    Article  CAS  PubMed  Google Scholar 

  2. Harris, A. L. Hypoxia — a key regulatory factor in tumour growth. Nature Rev. Cancer 2, 38–47 (2002).

    Article  CAS  Google Scholar 

  3. Semenza, G. L. HIF-1 and tumor progression: pathophysiology and therapeutics. Trends Mol. Med. 8, S62–S67 (2002).

    Article  CAS  PubMed  Google Scholar 

  4. Thompson, E. W., Newgreen, D. F. & Tarin, D. Carcinoma invasion and metastasis: a role for epithelial–mesenchymal transition? Cancer Res. 65, 5991–5995 (2005).

    Article  CAS  PubMed  Google Scholar 

  5. Thiery, J. P. Epithelial-mesenchymal transitions in tumour progression. Nature Rev. Cancer 2, 442–454 (2002).

    Article  CAS  Google Scholar 

  6. Thiery, J. P. & Sleeman, J. P. Complex networks orchestrate epithelial-mesenchymal transitions. Nature Rev. Mol. Cell Biol. 7, 131–142 (2006).

    Article  CAS  Google Scholar 

  7. Batlle, E. et al. The transcription factor Snail is a repressor of E-cadherin gene expression in epithelial tumour cells. Nature Cell Biol. 2, 84–89 (2000).

    Article  CAS  PubMed  Google Scholar 

  8. Cano, A. et al. The transcription factor Snail controls epithelial-mesenchymal transitions by repressing E-cadherin expression. Nature Cell Biol. 2, 76–83 (2000).

    Article  CAS  PubMed  Google Scholar 

  9. Peinado, H., Olmedo, D. & Cano, A. Snail, ZEB, and bHLH factors in tumour progression: an alliance against the epithelial phenotype? Nature Rev. Cancer 7, 415–428 (2007).

    Article  CAS  Google Scholar 

  10. Moody, S. E. et al. The transcriptional repressor Snail promotes mammary tumor recurrence. Cancer Cell 8, 197–209 (2005).

    Article  CAS  PubMed  Google Scholar 

  11. Imai, T. et al. Hypoxia attenuates the expression of E-cadherin via up-regulation of SNAIL in ovarian carcinoma cells. Am. J. Pathol. 161, 1437–1447 (2003).

    Article  Google Scholar 

  12. Krishnamachary, B. et al. Hypoxia-inducible factor-1-dependent repression of E-cadherin in von Hippel-Lindau tumor suppressor-null renal carcinoma mediated by TCF3, ZFHX1A, and ZFHX1B. Cancer Res. 66, 2725–2731 (2006).

    Article  CAS  PubMed  Google Scholar 

  13. Evans, A. J. et al. VHL promotes E2 box-dependent E-cadherin transcription by HIF-mediated regulation of SIP1 and Snail. Mol. Cell. Biol. 27, 157–169 (2007).

    Article  CAS  PubMed  Google Scholar 

  14. Erler, J.T., et al. Lysyl oxidase is essential for hypoxia-induced metastasis. Nature 440, 1222–1226 (2006).

    Article  CAS  PubMed  Google Scholar 

  15. Castanon, I. & Baylies, M. K. Twist in fate: evolutionary comparison of Twist structure and function. Gene 287, 11–22 (2002).

    Article  CAS  PubMed  Google Scholar 

  16. Furlong, E. E., Andersen, E. C., Null, B., White, K. P. & Scott, M. P. Patterns of gene expression during Drosophila mesoderm development. Science 293, 1629–1633 (2001).

    Article  CAS  PubMed  Google Scholar 

  17. Yang, J., et al. Twist, a master regulator of morphogenesis, plays an essential role in tumor metastasis. Cell 117, 927–939 (2004).

    Article  CAS  Google Scholar 

  18. Lee, T. K., et al. Twist overexpression correlates with hepatocellular carcinoma metastasis through induction of epithelial-mesenchymal transition. Clin. Cancer Res. 12, 5369–5376 (2006).

    Article  CAS  PubMed  Google Scholar 

  19. Ryan, H. E. Lo, J., & Johnson, R. S. HIF-1α is required for solid tumor formation and embryonic vascularization. EMBO J. 17, 3005–3115 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Iyer, N.V. et al. Cellular and developmental control of O2 homeostasis by hypoxia-inducible factor 1α. Genes Dev. 12, 149–162 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Chen, Z. F. & Behringer, R. R. Twist is required in head mesenchyme for cranial neural tube morphogenesis. Genes Dev. 9, 686–699 (1995).

    Article  CAS  PubMed  Google Scholar 

  22. Krishnamachary, B., et al. Regulation of colon carcinoma cell invasion by hypoxia-inducible factor 1. Cancer Res. 63, 1138–43 (2003).

    CAS  PubMed  Google Scholar 

  23. Huang, L. E., Gu, J., Schau, M. & Bunn, H. F. Regulation of hypoxia-inducible factor 1α is mediated by an O2-dependent degradation domain via the ubiquitin-proteasome pathway. Proc. Natl Acad. Sci. USA 95, 7987–7992 (1998).

    Article  CAS  PubMed  Google Scholar 

  24. Sawair, F. A., et al. Invasive front grading: reliability and usefulness in the management of oral squamous cell carcinoma. J. Oral Pathol. Med. 32, 1–9 (2003).

    Article  PubMed  Google Scholar 

  25. Hirota, K. & Semenza, G. L. Regulation of angiogenesis by hypoxia-inducible factor 1. Crit. Rev. Oncol. Hematol. 59, 15–26 (2006).

    Article  PubMed  Google Scholar 

  26. Alexander, N.R., et al. N-cadherin gene expression in prostate carcinoma is modulated by integrin-dependent nuclear translocation of Twist1. Cancer Res. 66, 3365–3369 (2006).

    Article  CAS  PubMed  Google Scholar 

  27. Koshiji, M., et al. HIF-1α induces cell cycle arrest by functionally counteracting Myc. EMBO J. 23, 1949–1956 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Janssen, H. L., Haustermans, K. M., Balm, A. J. & Begg, A. C. Hypoxia in head and neck cancer: how much, how important? Head Neck 27, 622–638 (2005).

    Article  CAS  PubMed  Google Scholar 

  29. Pugh, C. W., Gleadle, J. & Maxwell, P.H. Hypoxia and oxidative stress in breast cancer: hypoxia signalling pathways. Breast Cancer Res. 3, 313–317 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Yang, M.H., et al. Overexpression of NBS1 induces epithelial-mesenchymal transition and co-expression of NBS1 and Snail predicts metastasis of head and neck cancer. Oncogene 26, 1459–1467 (2007).

    Article  CAS  PubMed  Google Scholar 

  31. Koukourakis, M.I., et al. Endogenous markers of two separate hypoxia response pathways (hypoxia-inducible factor 2α and carbonic anhydrase 9) are associated with radiotherapy failure in head and neck cancer patients recruited in the CHART randomized trial. J. Clin. Oncol. 24, 727–735 (2006).

    Article  CAS  PubMed  Google Scholar 

  32. Hui, E. P., et al. Coexpression of hypoxia-inducible factors 1α and 2α, carbonic anhydrase IX and vascular endothelial growth factor in nasopharyngeal carcinoma and relationship to survival. Clin. Cancer Res. 8, 2595–2604 (2002).

    CAS  PubMed  Google Scholar 

  33. Trastour, C., et al. HIF-1α and CA IX staining in invasive breast carcinomas: prognosis and treatment outcome. Int. J. Cancer 120, 1451–1458 (2007).

    Article  CAS  PubMed  Google Scholar 

  34. Corsi, A. K., Kostas, S. A., Fire, A. & Krause, M. Caenorhabditis elegans Twist plays an essential role in non-striated muscle development. Development 127, 2041–2051 (2000).

    CAS  PubMed  Google Scholar 

  35. Provot, S. et al. Hif-1α regulates differentiation of limb bud mesenchyme and joint development. J. Cell Biol. 177, 451–464 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Wu, K. J., et al. Direct activation of TERT transcription by c-Myc. Nature Genet. 21, 220–224 (1999).

    Article  CAS  PubMed  Google Scholar 

  37. Chiang, Y. C., Teng, S. C., Su, Y. N., Hsieh, F. J. & Wu, K. J. c-Myc directly regulates the transcription of the NBS1 gene involved in DNA double-strand break repair. J. Biol. Chem. 278, 19286–19291 (2003).

    Article  CAS  PubMed  Google Scholar 

  38. Tickoo, S. K., et al. Immunohistochemical expression of hypoxia-inducible factor-1α and its downstream molecules in sarcomatoid renal cell carcinoma. J. Urol. 177, 1258–1263 (2007).

    Article  CAS  PubMed  Google Scholar 

  39. Maestro, R., et al. Twist is a potential oncogene that inhibits apoptosis. Genes Dev. 13, 2207–2217 (1999).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank K. W. Chang for TMA construction, and C. H. Huang and D. H. Hsu for excellent technical assistance. We are grateful to T. Y. Chou and W. Y. Li for providing expert opinions on pathology reading and immunohistochemical analysis. This work was supported in part by National Research Program for Genomic Medicine (DOH-96-TD-G-111-002)(K.J.W.), Taipei Veterans General Hospital VGH 96-C1-126, V-96-ER2-008 (M.H.Y.), V-96-ER2-009 (S.Y.C.), National Science Council (NSC-95-2320-B-010-065)(K.J.W.); (95-2314-B-075-083)(M.H.Y.), a grant from Ministry of Education, Aim for the Top University Plan (95A-C-D01-PPG-05, 96A-D-D139)(K.J.W.), and National Health Research Institutes (NHRI-EX-96-9611BI)(K.J.W.).

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Authors

Contributions

K. J. W., M. H. Y. and M. Z. W. conceived and designed the experiments; M. H. Y. and M. Z. W. performed the experiments with the assistance of S. H. C. for in vivo work; M. H. Y., M. Z. W., S. C. T. and K. J. W. analysed the data; K. J. W. and M. H. Y. wrote the paper with the assistance of S. C. T; S. Y. C., C. J. L., P. M. C. and M. H. Y. collected and treated samples from head and neck cancer patients.

Corresponding author

Correspondence to Kou-Juey Wu.

Supplementary information

Supplementary Information

Supplementary figures S1, S2, S3, S4, S5, S6, Supplementary tables S1, S2, S3, S4, S5, S6 and Supplementary Methods (PDF 1824 kb)

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Yang, MH., Wu, MZ., Chiou, SH. et al. Direct regulation of TWIST by HIF-1α promotes metastasis. Nat Cell Biol 10, 295–305 (2008). https://doi.org/10.1038/ncb1691

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