Annals of Surgical Oncology

, Volume 12, Issue 6, pp 488–496 | Cite as

Expression of the Transcription Factors Snail, Slug, and Twist and Their Clinical Significance in Human Breast Cancer

  • Tracey A. Martin
  • Amit Goyal
  • Gareth Watkins
  • Wen G. Jiang
Article

Abstract

Background

Slug, Snail, and Twist are transcription factors that regulate the expression of tumor suppressors such as E-cadherin. We examined the distribution and expression of these three molecules together with the methylation of the Twist gene promoter in human breast cancer to elucidate their clinical significance.

Methods

Frozen sections from breast cancer primary tumors (tumor, n = 114; background, n = 30) were immunostained with Slug, Snail, and Twist antibodies. RNA was reverse-transcribed, quantified, and analyzed by quantitative polymerase chain reaction (Q-PCR). Results were expressed as copy number of transcript per 50 ng of RNA (standardized against β-actin).

Results

Immunohistochemistry revealed that all three molecules were stained in mammary tissues, with an increase in Twist within tumor tissues; this was supported by Q-PCR analysis. Q-PCR analysis showed that Slug was elevated with increasing tumor grade and prognostic indices. Twist was elevated with increasing nodal involvement (tumor-node-metastasis status). Conversely, Snail was reduced in expression corresponding with prognostic indices and tumor grade. Increased levels of Slug were associated with tumors from patients with metastatic disease or disease recurrence, and increased expression of Twist was associated with tumors from patients who had died from breast cancer. It is interesting to note that Snail expression was significantly reduced in patients with a poor outcome and those who had node-positive tumors. In addition, tumors exhibited methylation of the Twist promoter.

Conclusions

These data demonstrate that all three transcription factors have inappropriate expression in breast cancer and that this may play a part in the progression of human breast tumors.

Keywords

Breast cancer Twist Snail Slug 

References

  1. 1.
    Seki, K, Fujimori, T, Savagner, P,  et al. 2003Mouse Snail family transcription repressors regulate chondrocyte, extracellular matrix, type II collagen, and aggrecanBiol Chem2784186270CrossRefGoogle Scholar
  2. 2.
    Boulay, JL, Dennefeld, C, Alberga, A 1987The Drosophila developmental gene snail encodes a protein with nucleic acid binding fingersNature3303958CrossRefPubMedGoogle Scholar
  3. 3.
    Hemavathy, K, Ashraf, SI, Ip, YT 2000Snail/slug family of repressors: slowly going into the fast lane of development and cancerGene257112CrossRefPubMedGoogle Scholar
  4. 4.
    Arias, AM 2001Epithelial mesenchymal interactions in cancer and developmentCell10542531CrossRefPubMedGoogle Scholar
  5. 5.
    Sugimachi, K, Tanaka, S, Kameyama, T,  et al. 2003Transcriptional repressor snail and progression of human hepatocellular carcinomaClin Cancer Res9265764PubMedGoogle Scholar
  6. 6.
    Hajra, KM, Chen, DY, Fearon, ER 2002The SLUG zinc-finger protein represses E-cadherin in breast cancerCancer Res6216138PubMedGoogle Scholar
  7. 7.
    Bolos, V, Peinado, H, Perez-Moreno, MA, Fraga, MF, Esteller, M, Cano, A 2003The transcription factor Slug represses E-cadherin expression and induces epithelial to mesenchymal transitions: a comparison with Snail and E47 repressorsJ Cell Sci116499511CrossRefPubMedGoogle Scholar
  8. 8.
    Jiang, WG 1996E-cadherin and its associated protein catenins, cancer invasion and metastasisBr J Surg8343746PubMedGoogle Scholar
  9. 9.
    Hamamori, Y, Wu, HY, Sartorelli, V, Kedes, L 1997The basic domain of myogenic basic helix-loop-helix (bHLH) proteins is the novel target for direct inhibition by another bHLH protein, TwistMol Cell Biol17656373PubMedGoogle Scholar
  10. 10.
    Maestro, R, Dei Tos, AP, Hamamori, Y,  et al. 1999Twist is a potential oncogene that inhibits apoptosisGenes Dev13220717CrossRefPubMedGoogle Scholar
  11. 11.
    Castanon, I, Stetina, S, Kass, J, Baylies, MK 2001Dimerization partners determine the activity of the Twist bHLH protein during Drosophila mesoderm developmentDevelopment128314559PubMedGoogle Scholar
  12. 12.
    Oda, H, Tsukita, S, Takeichi, M 1998Dynamic behavior of the cadherin-based cell-cell adhesion system during Drosophila gastrulationDev Biol20343550CrossRefPubMedGoogle Scholar
  13. 13.
    Villavicencio, EH, Yoon, JW, Frank, DJ, Fuchtbauer, EM, Walterhouse, DO, Iannaccone, PM 2002Cooperative E-box regulation of human GLI1 by TWIST and USFGenesis3224758CrossRefPubMedGoogle Scholar
  14. 14.
    Evron, E, Dooley, WC, Umbricht, CB,  et al. 2001Detection of breast cancer cells in ductal lavage fluid by methylation-specific PCRLancet35713356CrossRefPubMedGoogle Scholar
  15. 15.
    Wang, X, Ling, MT, Guan, XY,  et al. 2004Identification of a novel function of TWIST, a bHLH protein, in the development of acquired taxol resistance in human cancer cellsOncogene2347482CrossRefPubMedGoogle Scholar
  16. 16.
    Thiery, JP 2002Epithelial-mesenchymal transitions in tumour progressionNat Rev Genet244254Google Scholar
  17. 17.
    Boyer, B, Valles, AM, Edme, N 2000Induction and regulation of epithelial-mesenchymal transitionsBiochem Pharmacol6010919CrossRefPubMedGoogle Scholar
  18. 18.
    Jechlinger, M, Grunert, S, Tamir, IH,  et al. 2003Expression profiling of epithelial plasticity in tumor progressionOncogene22715569CrossRefPubMedGoogle Scholar
  19. 19.
    Thiery, JP 2003Epithelial-mesenchymal transitions in development and pathologiesCurr Opin Cell Biol157406CrossRefPubMedGoogle Scholar
  20. 20.
    Huber, MA, Azoitei, N, Baumann, B,  et al. 2004NF-κB is essential for epithelial-mesenchymal transition and metastasis in a model of breast cancer progressionJ Clin Invest11456981CrossRefPubMedGoogle Scholar
  21. 21.
    Kang, Y, Massague, J 2004Epithelial-mesenchymal transitions: twist in development and metastasisCell1182779CrossRefPubMedGoogle Scholar
  22. 22.
    Yang, J, Mani, SA, Donahar, JL,  et al. 2004Twist, a master regulator of morphogenesis, plays an essential role in tumor metastasisCell11892739CrossRefGoogle Scholar
  23. 23.
    Jiang, WG, Watkins, G, Lane, J,  et al. 2003Prognostic value of rho GTPases and rho guanine nucleotide dissociation inhibitors in human breast cancersClin Cancer Res9643240PubMedGoogle Scholar
  24. 24.
    Herman, JG, Graff, JR, Myohanen, S, Nelkin, BD, Baylin, SB 1996Methylation-specific PCR: a novel PCR assay for methylation status of CpG islandsProc Natl Acad Sci U S A9398216CrossRefPubMedGoogle Scholar
  25. 25.
    Nieto, MA, Sargent, MG, Wilkinson, DG, Cooke, J 1994Control of cell behavior during vertebrate development by Slug, a zinc finger geneScience2648359PubMedGoogle Scholar
  26. 26.
    Cano, A, Perez-Moreno, MA, Rodrigo, I,  et al. 2000The transcription factor snail controls epithelial-mesenchymal transitions by repressing E-cadherin expressionNat Cell Biol 27683CrossRefPubMedGoogle Scholar
  27. 27.
    Carver, EA, Jiang, R, Lan, Y, Oram, KF, Gridley, T 2001The mouse snail gene encodes a key regulator of the epithelial-mesenchymal transitionMol Cell Biol2181848CrossRefPubMedGoogle Scholar
  28. 28.
    Batlle, E, Sancho, E, Franci, C,  et al. 2000The transcription factor snail is a repressor of E-cadherin gene expression in epithelial tumor cellsNat Cell Biol2849CrossRefPubMedGoogle Scholar
  29. 29.
    Cheng, CW, Wu, PE, Yu, JC,  et al. 2001Mechanisms of inactivation of E-cadherin in breast carcinoma: modification of the two-hit hypothesis of tumor suppressor geneOncogene20381423CrossRefPubMedGoogle Scholar
  30. 30.
    Blanco, MJ, Moreno-Bueno, G, Sarrio, D,  et al. 2002Correlation of Snail expression with histological grade and lymph node status in breast carcinomasOncogene2132416CrossRefPubMedGoogle Scholar
  31. 31.
    Goyal, A, Douglas-Jones, A, Mansel, RE, Jiang, WG 2003E-cadherin and alpha-catenin expression in relation to clinical outcomes in patients with breast cancer. Presented to the 26th San Antonio Breast Cancer SymposiumBreast Cancer Res Treat82517Google Scholar
  32. 32.
    Okubo, T, Truong, TK, Yu, B,  et al. 2001Down-regulation of promoter 1.3 activity of the human aromatase gene in breast tissue by zinc-finger protein, snail (SnaH)Cancer Res61133846PubMedGoogle Scholar
  33. 33.
    Chen, S, Itoh, T, Wu, K, Zhou, D, Yang, C 2002Transcriptional regulation of aromatase expression in human breast tissueJ Steroid Biochem Mol Biol83939CrossRefPubMedGoogle Scholar
  34. 34.
    Chen, S, Zhou, D, Okubo, T,  et al. 2002Prevention and treatment of breast cancer by suppressing aromatase activity and expressionAnn N Y Acad Sci96322938PubMedCrossRefGoogle Scholar
  35. 35.
    Inukai, T, Inoue, A, Kurosawa, H,  et al. 1999SLUG, a ces-1-related zinc finger transcription factor gene with antiapoptotic activity, is a downstream target of the E2A-HLF oncoproteinMol Cell434352CrossRefPubMedGoogle Scholar
  36. 36.
    Poser, I, Dominguez, D, de Herreros, AG, Varnai, A, Buettner, R, Bosserhoff, AK 2001Loss of E-cadherin expression in melanoma cells involves up-regulation of the transcriptional repressor SnailJ Biol Chem276246616CrossRefPubMedGoogle Scholar
  37. 37.
    Aybar, MJ, Nieto, MA, Mayor, R 2003Snail precedes slug in the genetic cascade required for the specification and migration of the Xenopus neural crestDevelopment13048394CrossRefPubMedGoogle Scholar
  38. 38.
    Nieman, MT, Prudoff, RS, Johnson, KR, Wheelock, MJ 1999N-cadherin promotes motility in human breast cancer cells regardless of their E-cadherin expressionJ Cell Biol14763144CrossRefPubMedGoogle Scholar
  39. 39.
    Li, G, Satyamoorthy, K, Herlyn, M 2001N-cadherin-mediated intercellular interactions promote survival and migration of melanoma cellsCancer Res61381925PubMedGoogle Scholar
  40. 40.
    Rosivatz, E, Becker, I, Specht, K,  et al. 2002Differential expression of the epithelial-mesenchymal transition regulators snail, SIP1, and twist in gastric cancerAm J Pathol161188191PubMedGoogle Scholar
  41. 41.
    Fackler, MJ, McVeigh, M, Evron, E,  et al. 2003DNA methylation of RASSF1A, HIN-1, RAR-beta, cyclin D2 and Twist in in situ and invasive lobular breast carcinomaInt J Cancer1079705CrossRefPubMedGoogle Scholar
  42. 42.
    Howe, LR, Watanabe, O, Leonard, J, Brown, AM 2003Twist is up-regulated in response to Wnt1 and inhibits mouse mammary cell differentiationCancer Res63190613PubMedGoogle Scholar

Copyright information

© The Society of Surgical Oncology, Inc. 2005

Authors and Affiliations

  • Tracey A. Martin
    • 1
  • Amit Goyal
    • 1
  • Gareth Watkins
    • 1
  • Wen G. Jiang
    • 1
  1. 1.Metastasis & Angiogenesis Research Group, University Department of Surgery, Wales College of MedicineCardiff UniversityHeath ParkUnited Kingdom

Personalised recommendations