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The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1

Nature Cell Biology volume 10, pages 593–601 (2008)Cite this article

Abstract

Epithelial to mesenchymal transition (EMT) facilitates tissue remodelling during embryonic development and is viewed as an essential early step in tumour metastasis. We found that all five members of the microRNA-200 family (miR-200a, miR-200b, miR-200c, miR-141 and miR-429) and miR-205 were markedly downregulated in cells that had undergone EMT in response to transforming growth factor (TGF)-β or to ectopic expression of the protein tyrosine phosphatase Pez. Enforced expression of the miR-200 family alone was sufficient to prevent TGF-β-induced EMT. Together, these microRNAs cooperatively regulate expression of the E-cadherin transcriptional repressors ZEB1 (also known as δEF1) and SIP1 (also known as ZEB2), factors previously implicated in EMT and tumour metastasis. Inhibition of the microRNAs was sufficient to induce EMT in a process requiring upregulation of ZEB1 and/or SIP1. Conversely, ectopic expression of these microRNAs in mesenchymal cells initiated mesenchymal to epithelial transition (MET). Consistent with their role in regulating EMT, expression of these microRNAs was found to be lost in invasive breast cancer cell lines with mesenchymal phenotype. Expression of the miR-200 family was also lost in regions of metaplastic breast cancer specimens lacking E-cadherin. These data suggest that downregulation of the microRNAs may be an important step in tumour progression.

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Figure 1: Identification of microRNAs that are regulated during EMT.
Figure 2: The miRNA-200 family and miRNA-205 are downregulated during TGF-β-mediated EMT.
Figure 3: ZEB1 and SIP1 3´UTRs are regulated by miR-200a, miR-200b and miR-205.
Figure 4: Manipulation of the miR-200 family and miR-205 levels can induce EMT or MET.
Figure 5: MicroRNA levels are associated with mesenchymal phenotype.

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References

  1. Bagga, S. et al. Regulation by let-7 and lin-4 miRNAs results in target mRNA degradation. Cell 122, 553–563 (2005).

    Article  CAS  Google Scholar 

  2. Lim, L. P. et al. Microarray analysis shows that some micro RNAs downregulate large numbers of target mRNAs. Nature 433, 769–773 (2005).

    Article  CAS  Google Scholar 

  3. Fazi, F. et al. A minicircuitry comprised of microRNA-223 and transcription factors NFI-A and C/EBPα regulates human granulopoiesis. Cell 123, 819–831 (2005).

    Article  CAS  Google Scholar 

  4. Wyatt, L., Wadham, C., Crocker, L. A., Lardelli, M. & Khew-Goodall, Y. The protein tyrosine phosphatase Pez regulates TGF-β, epithelial mesenchymal transition, and organ development. J. Cell Biol. 178, 1223–1235 (2007).

    Article  CAS  Google Scholar 

  5. Lewis, B. P., Burge, C. B. & Bartel, D. P. Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 120, 15–20 (2005).

    Article  CAS  Google Scholar 

  6. Comijn, J. et al. The two-handed E box binding zinc finger protein SIP1 downregulates E-cadherin and induces invasion. Mol. Cell 7, 1267–1278 (2001).

    Article  CAS  Google Scholar 

  7. Eger, A. et al. δEF1 is a transcriptional repressor of E-cadherin and regulates epithelial plasticity in breast cancer cells. Oncogene 24, 2375–2385 (2005).

    Article  CAS  Google Scholar 

  8. Hurteau, G. J., Carlson, J. A., Spivack, S. D. & Brock, G. J. Overexpression of the microRNA hsa-miR-200c leads to reduced expression of transcription factor 8 and increased expression of E-cadherin. Cancer Res. 67, 7972–7976 (2007).

    Article  CAS  Google Scholar 

  9. Christoffersen, N. R., Silahtaroglu, A., Orom, U. A., Kauppinen, S. & Lund, A. H. miR-200b mediates post-transcriptional repression of ZFHX1B. RNA 13, 1172–1178 (2007).

    Article  CAS  Google Scholar 

  10. 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 

  11. Huber, M. A. et al. NF-κB is essential for epithelial-mesenchymal transition and metastasis in a model of breast cancer progression. J. Clin. Invest. 114, 569–581 (2004).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  13. Lacroix, M. & Leclercq, G. Relevance of breast cancer cell lines as models for breast tumours: an update. Breast Cancer Res. Treat. 83, 249–289 (2004).

    Article  CAS  Google Scholar 

  14. Park, S. M., Gaur, A. B., Lengyel, E. & Peter, M. E. The miR-200 family determines the epithelial phenotype of cancer cells by targeting the E-cadherin repressors, ZEB1 and ZEB2. Genes Dev. advance online publication, doi:10.1101/gad.1640608 (2008).

  15. Lien, H. C. et al. Molecular signatures of metaplastic carcinoma of the breast by large-scale transcriptional profiling: identification of genes potentially related to epithelial-mesenchymal transition. Oncogene 26, 7859–7871 (2007).

    Article  CAS  Google Scholar 

  16. Sempere, L. F. et al. Altered microRNA expression confined to specific epithelial cell subpopulations in breast cancer. Cancer Res. 67, 11612–11620 (2007).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  18. Thomson, J. M., Parker, J., Perou, C. M. & Hammond, S. M. A custom microarray platform for analysis of microRNA gene expression. Nature Methods 1, 47–53 (2004).

    Article  CAS  Google Scholar 

  19. Baskerville, S. & Bartel, D. P. Microarray profiling of microRNAs reveals frequent coexpression with neighboring miRNAs and host genes. RNA 11, 241–247 (2005).

    Article  CAS  Google Scholar 

  20. Lu, J. et al. MicroRNA expression profiles classify human cancers. Nature 435, 834–838 (2005).

    Article  CAS  Google Scholar 

  21. Wienholds, E. et al. MicroRNA expression in zebrafish embryonic development. Science 309, 310–311 (2005).

    Article  CAS  Google Scholar 

  22. Darnell, D. K. et al. Micro RNA expression during chick embryo development. Dev. Dyn. 235, 3156–3165 (2006).

    Article  CAS  Google Scholar 

  23. Funahashi, J., Sekido, R., Murai, K., Kamachi, Y. & Kondoh, H. δ-crystallin enhancer binding protein delta EF1 is a zinc finger-homeodomain protein implicated in postgastrulation embryogenesis. Development 119, 433–446 (1993).

    CAS  PubMed  Google Scholar 

  24. Miyoshi, T. et al. Complementary expression pattern of Zfhx1 genes Sip1 and δEF1 in the mouse embryo and their genetic interaction revealed by compound mutants. Dev. Dyn. 235, 1941–1952 (2006).

    Article  CAS  Google Scholar 

  25. Yi, R. et al. Morphogenesis in skin is governed by discrete sets of differentially expressed microRNAs. Nature Genet. 38, 356–362 (2006).

    Article  CAS  Google Scholar 

  26. 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 

  27. Peinado, H., Portillo, F. & Cano, A. Transcriptional regulation of cadherins during development and carcinogenesis. Int. J. Dev. Biol. 48, 365–375 (2004).

    Article  CAS  Google Scholar 

  28. Ohira, T. et al. WNT7a induces E-cadherin in lung cancer cells. Proc. Natl Acad. Sci. USA 100, 10429–10434 (2003).

    Article  CAS  Google Scholar 

  29. Spoelstra, N. S. et al. The transcription factor ZEB1 is aberrantly expressed in aggressive uterine cancers. Cancer Res. 66, 3893–3902 (2006).

    Article  CAS  Google Scholar 

  30. Spaderna, S. et al. A transient, EMT-linked loss of basement membranes indicates metastasis and poor survival in colorectal cancer. Gastroenterology 131, 830–840 (2006).

    Article  CAS  Google Scholar 

  31. Lombaerts, M. et al. E-cadherin transcriptional downregulation by promoter methylation but not mutation is related to epithelial-to-mesenchymal transition in breast cancer cell lines. Br. J. Cancer 94, 661–671 (2006).

    Article  CAS  Google Scholar 

  32. Raymond, C. K., Roberts, B. S., Garrett-Engele, P., Lim, L. P. & Johnson, J. M. Simple, quantitative primer-extension PCR assay for direct monitoring of microRNAs and short-interfering RNAs. RNA 11, 1737–1744 (2005).

    Article  CAS  Google Scholar 

  33. Smyth, G. K. Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Stat. Appl. Genet. Mol. Biol. 3, Article3 (2004).

  34. Barry, S. C. et al. Lentivirus vectors encoding both central polypurine tract and posttranscriptional regulatory element provide enhanced transduction and transgene expression. Hum. Gene Ther. 12, 1103–1108 (2001).

    Article  CAS  Google Scholar 

  35. Pillai, R. S. et al. Inhibition of translational initiation by Let-7 MicroRNA in human cells. Science 309, 1573–1576 (2005).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors thank Don Newgreen for critical reading of the manuscript, Witold Filipowicz for the pCI-neo-hRL and RL-let-7 plasmids, Scott Hammond for the pLenti4.1EX plasmid, Mark van der Hoek and the Adelaide Microarray Facility for printing of the microarrays, and members of the Goodall and Khew-Goodall laboratories for helpful discussions. This work was supported by grants from the National Health and Medical Research Council of Australia (to Y. K.-G., M. A. V. and G. J. G.) and The Cancer Council South Australia (to G. J. G.). P. A. G. is supported by a Peter Doherty Training Fellowship from the National Health and Medical Research Council of Australia.

Author information

Author notes

  1. Mathew A. Vadas

    Present address: Current address, Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Newtown, NSW 2042, Australia.,

  2. Yeesim Khew-Goodall and Gregory J. Goodall: These authors contributed equally to this study.

Authors and Affiliations

  1. Hanson Institute and Division of Human Immunology, Institute of Medical and Veterinary Science, Adelaide, 5000, SA, Australia

    Philip A. Gregory, Andrew G. Bert, Emily L. Paterson, Anna Tsykin, Mathew A. Vadas, Yeesim Khew-Goodall & Gregory J. Goodall

  2. Discipline of Paediatrics, School of Paediatrics and Reproductive Health, University of Adelaide, Adelaide, SA 5005

    Simon C. Barry

  3. Women's and Children's Health Research Institute, Adelaide, 5006, SA, Australia

    Simon C. Barry

  4. Division of Tissue Pathology, Institute of Medical and Veterinary Science, Adelaide, 5000, SA, Australia

    Gelareh Farshid

  5. Discipline of Medicine, The University of Adelaide, Adelaide, 5005, SA, Australia

    Philip A. Gregory, Mathew A. Vadas & Gregory J. Goodall

  6. School of Molecular and Biomedical Sciences, The University of Adelaide, Adelaide, 5005, SA, Australia

    Yeesim Khew-Goodall

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Supplementary Figures S1, S2, S3, S4, S5 and Supplementary Table 1 (PDF 1400 kb)

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Gregory, P., Bert, A., Paterson, E. et al. The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol 10, 593–601 (2008). https://doi.org/10.1038/ncb1722

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