Tumor Biology

, Volume 31, Issue 4, pp 297–307 | Cite as

Slug/SNAI2 regulates cell proliferation and invasiveness of metastatic prostate cancer cell lines

  • Modjtaba Emadi Baygi
  • Zahra-Soheila Soheili
  • Frank Essmann
  • Abdolkhaleg Deezagi
  • Rainer Engers
  • Wolfgang Goering
  • Wolfgang A. SchulzEmail author
Research Article


Many metastatic cancers recapitulate the epithelial-to-mesenchymal transition (EMT) resulting in enhanced cell motility and invasiveness. The EMT is regulated by several transcription factors, including the zinc finger protein SNAI2, also named Slug, which appears to exert additional functions during development and cancer progression. We have studied the function of SNAI2 in prostate cancer cells. Quantitative RT-PCR analysis showed strong SNAI2 expression particularly in the PC-3 and PC3-16 prostate carcinoma cell lines. Knockdown of SNAI2 by specific siRNA induced changes in EMT markers and inhibited invasion of both cell lines into a matrigel matrix. SNAI2 siRNA-treated cells did not tolerate detachment from the culture plates, likely at least in part due to downregulation of integrin α6β4. SNAI2 knockdown disturbed the microtubular and actin cytoskeletons, especially severely in PC-3 cells, resulting in grossly enlarged, flattened, and sometimes multinuclear cells. Knockdown also decreased cell proliferation, with a prominent G0/G1 arrest in PC3-16. Together, our data imply that SNAI2 exerts strong effects on the cytoskeleton and adhesion of those prostate cancer cells that express it and is necessary for their proliferation and invasiveness.


Slug Cell cycle arrest Cell shape Invasiveness Integrin α6β4 Cytoskeleton Epithelial–mesenchymal transition 



We thank Christiane Hader and Dr. Parvaneh Nikpour for valuable assistance in several experiments and helpful suggestions.

Supplementary material

13277_2010_37_MOESM1_ESM.doc (70 kb)
Supplementary Table 1 (DOC 70 kb)
13277_2010_37_Fig9_ESM.gif (2.2 mb)
Supplementary 1

DAPI staining of attached or floating PC-3 or PC3-16 cells after treatment with the indicated siRNAs. In these representative figures, no apoptotic nuclei are visible except one marked by an arrow. In particular, cells centrifuged down from the supernatant onto microscope slides appear clumpy, but not apoptotic. (GIF 2252 kb)

13277_2010_37_MOESM2_ESM.tif (12.4 mb)
High resolution image (TIFF 12706 kb)


  1. 1.
    Barrallo-Gimeno A, Nieto MA. The snail genes as inducers of cell movement and survival: implications in development and cancer. Development. 2005;132:3151–61.CrossRefPubMedGoogle Scholar
  2. 2.
    Baum B, Settleman J, Quinlan MP. Transitions between epithelial and mesenchymal states in development and disease. Semin Cell Dev Biol. 2008;19:294–308.CrossRefPubMedGoogle Scholar
  3. 3.
    Hemavathy K, Guru SC, Harris J, Chen JD, Ip YT. Human slug is a repressor that localizes to sites of active transcription. Mol Cell Biol. 2000;20:5087–95.CrossRefPubMedGoogle Scholar
  4. 4.
    Alves CC, Carneiro F, Hoefler H, Becker KF. Role of the epithelial-mesenchymal transition regulator slug in primary human cancers. Front Biosci. 2009;14:3035–50.CrossRefPubMedGoogle Scholar
  5. 5.
    Perez-Caro M, Bermejo-Rodriguez C, Gonzalez-Herrero I, Sanchez-Beato M, Piris MA, Sanchez-Garcia I. Transcriptomal profiling of the cellular response to DNA damage mediated by slug (snai2). Brit J Cancer. 2008;98:480–8.CrossRefPubMedGoogle Scholar
  6. 6.
    Bolos V, Peinado H, Perez-Moreno MA, Fraga MF, Esteller M, Cano A. The transcription factor slug represses e-cadherin expression and induces epithelial to mesenchymal transitions: a comparison with snail and e47 repressors. J Cell Sci. 2003;116:499–511.CrossRefPubMedGoogle Scholar
  7. 7.
    Hajra KM, Chen DY, Fearon ER. The slug zinc-finger protein represses e-cadherin in breast cancer. Cancer Res. 2002;62:1613–18.PubMedGoogle Scholar
  8. 8.
    Savagner P, Yamada KM, Thiery JP. The zinc-finger protein slug causes desmosome dissociation, an initial and necessary step for growth factor-induced epithelial-mesenchymal transition. J Cell Biol. 1997;137:1403–19.CrossRefPubMedGoogle Scholar
  9. 9.
    Tripathi MK, Misra S, Chaudhuri G. Negative regulation of the expressions of cytokeratins 8 and 19 by slug repressor protein in human breast cells. Biochem Biophys Res Commun. 2005;329:508–15.CrossRefPubMedGoogle Scholar
  10. 10.
    Perez-Mancera PA, Gonzalez-Herrero I, Maclean K, Turner AM, Yip MY, Sanchez-Martin M, et al. Slug (Snai2) overexpression in embryonic development. Cytogenet Genome Res. 2006;114:24–9.CrossRefPubMedGoogle Scholar
  11. 11.
    Perez-Mancera PA, Gonzalez-Herrero I, Perez-Caro M, Gutierrez-Cianca N, Flores T, Gutierrez-Adan A, et al. Slug in cancer development. Oncogene. 2005;24:3073–82.CrossRefPubMedGoogle Scholar
  12. 12.
    Lensch R, Gotz C, Andres C, Bex A, Lehmann J, Zwergel T, et al. Comprehensive genotypic analysis of human prostate cancer cell lines and sublines derived from metastases after orthotopic implantation in nude mice. Int J Oncol. 2002;21:695–706.PubMedGoogle Scholar
  13. 13.
    Wlazlinski A, Engers R, Hoffmann MJ, Hader C, Jung V, Muller M, et al. Downregulation of several fibulin genes in prostate cancer. Prostate. 2007;67:1770–80.CrossRefPubMedGoogle Scholar
  14. 14.
    Jafarnejad SM, Mowla SJ, Matin MM. Knocking-down the expression of nucleostemin significantly decreases rate of proliferation of rat bone marrow stromal stem cells in an apparently p53-independent manner. Cell Prolif. 2008;41:28–35.PubMedGoogle Scholar
  15. 15.
    Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, et al. A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci USA. 1995;92:9363–7.CrossRefPubMedGoogle Scholar
  16. 16.
    Janssen K, Pohlmann S, Janicke RU, Schulze-Osthoff K, Fischer U. Apaf-1 and caspase-9 deficiency prevents apoptosis in a bax-controlled pathway and promotes clonogenic survival during paclitaxel treatment. Blood. 2007;110:3662–72.CrossRefPubMedGoogle Scholar
  17. 17.
    Engers R, Springer E, Michiels F, Collard JG, Gabbert HE. Rac affects invasion of human renal cell carcinomas by up-regulating tissue inhibitor of metalloproteinases (Timp)-1 and Timp-2 expression. J Biol Chem. 2001;276:41889–97.CrossRefPubMedGoogle Scholar
  18. 18.
    Guarino M, Rubino B, Ballabio G. The role of epithelial-mesenchymal transition in cancer pathology. Pathology. 2007;39:305–18.CrossRefPubMedGoogle Scholar
  19. 19.
    Lee JM, Dedhar S, Kalluri R, Thompson EW. The epithelial-mesenchymal transition: new insights in signaling, development, and disease. J Cell Biol. 2006;172:973–81.CrossRefPubMedGoogle Scholar
  20. 20.
    Pouliot N, Connolly LM, Moritz RL, Simpson RJ, Burgess AW. Colon cancer cells adhesion and spreading on autocrine laminin-10 is mediated by multiple integrin receptors and modulated by egf receptor stimulation. Exp Cell Res. 2000;261:360–71.CrossRefPubMedGoogle Scholar
  21. 21.
    Ridley AJ, Schwartz MA, Burridge K, Firtel RA, Ginsberg MH, Borisy G, et al. Cell migration: integrating signals from front to back. Science. 2003;302:1704–9.CrossRefPubMedGoogle Scholar
  22. 22.
    Lipscomb EA, Mercurio AM. Mobilization and activation of a signaling competent alpha6beta4integrin underlies its contribution to carcinoma progression. Cancer Metastasis Rev. 2005;24:413–23.CrossRefPubMedGoogle Scholar
  23. 23.
    Kato T, Katabami K, Takatsuki H, Han SA, Takeuchi K, Irimura T, et al. Characterization of the promoter for the mouse alpha 3 integrin gene. Eur J Biochem. 2002;269:4524–32.CrossRefPubMedGoogle Scholar
  24. 24.
    Lin CS, Chen Y, Huynh T, Kramer R. Identification of the human alpha6 integrin gene promoter. DNA Cell Biol. 1997;16:929–37.CrossRefPubMedGoogle Scholar
  25. 25.
    Takaoka AS, Yamada T, Gotoh M, Kanai Y, Imai K, Hirohashi S. Cloning and characterization of the human beta4-integrin gene promoter and enhancers. J Biol Chem. 1998;273:33848–55.CrossRefPubMedGoogle Scholar
  26. 26.
    Turner FE, Broad S, Khanim FL, Jeanes A, Talma S, Hughes S, et al. Slug regulates integrin expression and cell proliferation in human epidermal keratinocytes. J Biol Chem. 2006;281:21321–31.CrossRefPubMedGoogle Scholar
  27. 27.
    Goode BL, Drubin DG, Barnes G. Functional cooperation between the microtubule and actin cytoskeletons. Curr Opin Cell Biol. 2000;12:63–71.CrossRefPubMedGoogle Scholar
  28. 28.
    Vannini I, Bonafe M, Tesei A, Rosetti M, Fabbri F, Storci G, et al. Short interfering rna directed against the slug gene increases cell death induction in human melanoma cell lines exposed to cisplatin and fotemustine. Cell Oncol. 2007;29:279–87.PubMedGoogle Scholar
  29. 29.
    Newkirk KM, MacKenzie DA, Bakaletz AP, Hudson LG, Kusewitt DF. Microarray analysis demonstrates a role for slug in epidermal homeostasis. J Invest Dermatol. 2008;128:361–9.CrossRefPubMedGoogle Scholar
  30. 30.
    Robson EJ, Khaled WT, Abell K, Watson CJ. Epithelial-to-mesenchymal transition confers resistance to apoptosis in three murine mammary epithelial cell lines. Differentiation. 2006;74:254–64.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2010

Authors and Affiliations

  • Modjtaba Emadi Baygi
    • 1
  • Zahra-Soheila Soheili
    • 2
  • Frank Essmann
    • 3
  • Abdolkhaleg Deezagi
    • 2
  • Rainer Engers
    • 4
  • Wolfgang Goering
    • 5
  • Wolfgang A. Schulz
    • 5
    Email author
  1. 1.Department of Genetics, Faculty of Basic SciencesTarbiat Modares UniversityTehranIran
  2. 2.Department of BiochemistryNational Institute of Genetic Engineering and BiotechnologyTehranIran
  3. 3.Interfaculty Institute of BiochemistryEberhard Karls UniversityTübingenGermany
  4. 4.Institute of PathologyHeinrich Heine UniversityDüsseldorfGermany
  5. 5.Department of UrologyHeinrich Heine UniversityDüsseldorfGermany

Personalised recommendations