Skip to main content

Advertisement

SpringerLink
Log in

Snail regulates cell survival and inhibits cellular senescence in human metastatic prostate cancer cell lines

  • Published:
Cell Biology and Toxicology Aims and scope Submit manuscript

Abstract

The epithelial–mesenchymal transition (EMT) is regarded as an important step in cancer metastasis. Snail, a master regulator of EMT, has been recently proposed to act additionally as a cell survival factor and inducer of motility. We have investigated the function of Snail (SNAI1) in prostate cancer cells by downregulating its expression via short (21-mer) interfering RNA (siRNA) and measuring the consequences on EMT markers, cell viability, death, cell cycle, senescence, attachment, and invasivity. Of eight carcinoma cell lines, the prostate carcinoma cell lines LNCaP and PC-3 showed the highest and moderate expression of SNAI1 mRNA, respectively, as measured by quantitative RT-PCR. Long-term knockdown of Snail induced a severe decline in cell numbers in LNCaP and PC-3 and caspase activity was accordingly enhanced in both cell lines. In addition, suppression of Snail expression induced senescence in LNCaP cells. SNAI1-siRNA-treated cells did not tolerate detachment from the extracellular matrix, probably due to downregulation of integrin α6. Expression of E-cadherin, vimentin, and fibronectin was also affected. Invasiveness of PC-3 cells was not significantly diminished by Snail knockdown. Our data suggest that Snail acts primarily as a survival factor and inhibitor of cellular senescence in prostate cancer cell lines. We therefore propose that Snail can act as early driver of prostate cancer progression.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11

Similar content being viewed by others

References

  • Ansieau S, Bastid J, Doreau A, Morel AP, Bouchet BP, Thomas C, et al. Induction of EMT by twist proteins as a collateral effect of tumor-promoting inactivation of premature senescence. Cancer Cell. 2008;14:79–89.

    Article  CAS  PubMed  Google Scholar 

  • Aybar MJ, Nieto MA, Mayor R. Snail precedes slug in the genetic cascade required for the specification and migration of the Xenopus neural crest. Development. 2003;130:483–94.

    Article  CAS  PubMed  Google Scholar 

  • Baritaki S, Chapman A, Yeung K, Spandidos DA, Palladino M, Bonavida B. Inhibition of epithelial to mesenchymal transition in metastatic prostate cancer cells by the novel proteasome inhibitor, NPI-0052: pivotal roles of Snail repression and RKIP induction. Oncogene. 2009;28:3573–85.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • Becker KF, Rosivatz E, Blechschmidt K, Kremmer E, Sarbia M, Hofler H. Analysis of the E-cadherin repressor Snail in primary human cancers. Cells Tissues Organs. 2007;185:204–12.

    Article  CAS  PubMed  Google Scholar 

  • Bhatia B, Jiang M, Suraneni M, Patrawala L, Badeaux M, Schneider-Broussard R, et al. Critical and distinct roles of p16 and telomerase in regulating the proliferative life span of normal human prostate epithelial progenitor cells. J Biol Chem. 2008;283:27957–72.

    Article  CAS  PubMed  Google Scholar 

  • Blanco MJ, Moreno-Bueno G, Sarrio D, Locascio A, Cano A, Palacios J, et al. Correlation of Snail expression with histological grade and lymph node status in breast carcinomas. Oncogene. 2002;21:3241–6.

    Article  CAS  PubMed  Google Scholar 

  • Blechschmidt K, Kremmer E, Hollweck R, Mylonas I, Hofler H, Kremer M, et al. The E-cadherin repressor snail plays a role in tumor progression of endometrioid adenocarcinomas. Diagn Mol Pathol. 2007;16:222–8.

    Article  CAS  PubMed  Google Scholar 

  • Borradori L, Sonnenberg A. Structure and function of hemidesmosomes: more than simple adhesion complexes. J Invest Dermatol. 1999;112:411–8.

    Article  CAS  PubMed  Google Scholar 

  • Caino MC, Meshki J, Kazanietz MG. Hallmarks for senescence in carcinogenesis: novel signaling players. Apoptosis. 2009;14:392–408.

    Article  CAS  PubMed  Google Scholar 

  • Castro P, Giri D, Lamb D, Ittmann M. Cellular senescence in the pathogenesis of benign prostatic hyperplasia. Prostate. 2003;55:30–8.

    Article  CAS  PubMed  Google Scholar 

  • Cervella P, Silengo L, Pastore C, Altruda F. Human beta 1-integrin gene expression is regulated by two promoter regions. J Biol Chem. 1993;268:5148–55.

    CAS  PubMed  Google Scholar 

  • Choi J, Shendrik I, Peacocke M, Peehl D, Buttyan R, Ikeguchi EF, et al. Expression of senescence-associated beta-galactosidase in enlarged prostates from men with benign prostatic hyperplasia. Urology. 2000;56:160–6.

    Article  CAS  PubMed  Google Scholar 

  • 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 U S A. 1995;92:9363–7.

    Article  CAS  PubMed  Google Scholar 

  • Dowling J, Yu QC, Fuchs E. Beta4 integrin is required for hemidesmosome formation, cell adhesion and cell survival. J Cell Biol. 1996;134:559–72.

    Article  CAS  PubMed  Google Scholar 

  • Edlund M, Miyamoto T, Sikes RA, Ogle R, Laurie GW, Farach-Carson MC, et al. Integrin expression and usage by prostate cancer cell lines on laminin substrata. Cell Growth Differ. 2001;12:99–107.

    CAS  PubMed  Google Scholar 

  • Elloul S, Elstrand MB, Nesland JM, Trope CG, Kvalheim G, Goldberg I, et al. Snail, Slug, and Smad-interacting protein 1 as novel parameters of disease aggressiveness in metastatic ovarian and breast carcinoma. Cancer. 2005;103:1631–43.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • Green KJ, Jones JC. Desmosomes and hemidesmosomes: structure and function of molecular components. Faseb J. 1996;10:871–81.

    CAS  PubMed  Google Scholar 

  • Guarino M, Rubino B, Ballabio G. The role of epithelial–mesenchymal transition in cancer pathology. Pathology. 2007;39:305–18.

    Article  CAS  PubMed  Google Scholar 

  • Haraguchi M, Okubo T, Miyashita Y, Miyamoto Y, Hayashi M, Crotti TN, et al. Snail regulates cell-matrix adhesion by regulation of the expression of integrins and basement membrane proteins. J Biol Chem. 2008;283:23514–23.

    Article  CAS  PubMed  Google Scholar 

  • Hardy RG, Vicente-Duenas C, Gonzalez-Herrero I, Anderson C, Flores T, Hughes S, et al. Snail family transcription factors are implicated in thyroid carcinogenesis. Am J Pathol. 2007;171:1037–46.

    Article  CAS  PubMed  Google Scholar 

  • Heebøll S, Borre M, Ottosen PD, Dyrskjøt L, Orntoft TF, Tørring N. Snail1 is over-expressed in prostate cancer. APMIS. 2009;117:196–204.

    Article  PubMed  Google Scholar 

  • Hornsby P. Senescence as an anticancer mechanism. J Clin Oncol. 2007;25:1852–7.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • Jarrard DF, Sarkar S, Shi Y, Yeager TR, Magrane G, Kinoshita H, et al. p16/pRb pathway alterations are required for bypassing senescence in human prostate epithelial cells. Cancer Res. 1999;59:2957–64.

    CAS  PubMed  Google Scholar 

  • Jemal A, Siegel R, Ward E, Murray T, Xu J, Thun MJ. Cancer statistics, 2007. CA Cancer J Clin. 2007;57:43–66.

    Article  PubMed  Google Scholar 

  • Kajita M, McClinic KN, Wade PA. Aberrant expression of the transcription factors snail and slug alters the response to genotoxic stress. Mol Cell Biol. 2004;24:7559–66.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • Kikkawa Y, Sanzen N, Fujiwara H, Sonnenberg A, Sekiguchi K. Integrin binding specificity of laminin-10/11: laminin-10/11 are recognized by alpha 3 beta 1, alpha 6 beta 1 and alpha 6 beta 4 integrins. J Cell Sci. 2000;113(Pt 5):869–76.

    CAS  PubMed  Google Scholar 

  • Lawrence MG, Veveris-Lowe TL, Whitbread AK, Nicol DL, Clements JA. Epithelial–mesenchymal transition in prostate cancer and the potential role of kallikrein serine proteases. Cells Tissues Organs. 2007;185:111–5.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    CAS  PubMed  Google Scholar 

  • Lin CS, Chen Y, Huynh T, Kramer R. Identification of the human alpha6 integrin gene promoter. DNA Cell Biol. 1997;16:929–37.

    Article  CAS  PubMed  Google Scholar 

  • Liu Y, El-Naggar S, Darling DS, Higashi Y, Dean DC. Zeb1 links epithelial–mesenchymal transition and cellular senescence. Development. 2008;135:579–88.

    Article  CAS  PubMed  Google Scholar 

  • Lukas J, Parry D, Aagaard L, Mann DJ, Bartkova J, Strauss M, et al. Retinoblastoma-protein-dependent cell-cycle inhibition by the tumour suppressor p16. Nature. 1995;375:503–6.

    Article  CAS  PubMed  Google Scholar 

  • Mercurio AM. Laminin receptors: achieving specificity through cooperation. Trends Cell Biol. 1995;5:419–23.

    Article  CAS  PubMed  Google Scholar 

  • Miyoshi A, Kitajima Y, Kido S, Shimonishi T, Matsuyama S, Kitahara K, et al. Snail accelerates cancer invasion by upregulating MMP expression and is associated with poor prognosis of hepatocellular carcinoma. Br J Cancer. 2005;92:252–8.

    CAS  PubMed  Google Scholar 

  • Nieto MA. The snail superfamily of zinc-finger transcription factors. Nat Rev Mol Cell Biol. 2002;3:155–66.

    Article  CAS  PubMed  Google Scholar 

  • Nishida K, Kitazawa R, Mizuno K, Maeda S, Kitazawa S. Identification of regulatory elements of human alpha 6 integrin subunit gene. Biochem Biophys Res Commun. 1997;241:258–63.

    Article  CAS  PubMed  Google Scholar 

  • Ohtani N, Mann DJ, Hara E. Cellular senescence: its role in tumor suppression and aging. Cancer Sci. 2009;100:792–7.

    Article  CAS  PubMed  Google Scholar 

  • Olmeda D, Jorda M, Peinado H, Fabra A, Cano A. Snail silencing effectively suppresses tumour growth and invasiveness. Oncogene. 2007a;26:1862–74.

    Article  CAS  PubMed  Google Scholar 

  • Olmeda D, Moreno-Bueno G, Flores JM, Fabra A, Portillo F, Cano A. SNAI1 is required for tumor growth and lymph node metastasis of human breast carcinoma MDA-MB-231 cells. Cancer Res. 2007b;67:11721–31.

    Article  CAS  PubMed  Google Scholar 

  • Olmeda D, Montes A, Moreno-Bueno G, Flores JM, Portillo F, Cano A. Snai1 and Snai2 collaborate on tumor growth and metastasis properties of mouse skin carcinoma cell lines. Oncogene. 2008;27:4690–701.

    Article  CAS  PubMed  Google Scholar 

  • Peinado H, Portillo F, Cano A. Transcriptional regulation of cadherins during development and carcinogenesis. Int J Dev Biol. 2004;48:365–75.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • Roy HK, Smyrk TC, Koetsier J, Victor TA, Wali RK. The transcriptional repressor SNAIL is overexpressed in human colon cancer. Dig Dis Sci. 2005;50:42–6.

    Article  CAS  PubMed  Google Scholar 

  • Sandhu C, Peehl DM, Slingerland J. p16INK4A mediates cyclin dependent kinase 4 and 6 inhibition in senescent prostatic epithelial cells. Cancer Res. 2000;60:2616–22.

    CAS  PubMed  Google Scholar 

  • Scher HI, Heller G. Clinical states in prostate cancer: toward a dynamic model of disease progression. Urology. 2000;55:323–7.

    Article  CAS  PubMed  Google Scholar 

  • Smit MA, Peeper DS. Deregulating EMT and senescence: double impact by a single twist. Cancer Cell. 2008;14:5–7.

    Article  CAS  PubMed  Google Scholar 

  • Stewart DA, Cooper CR, Sikes RA. Changes in extracellular matrix (ECM) and ECM-associated proteins in the metastatic progression of prostate cancer. Reprod Biol Endocrinol. 2004;2:2.

    Article  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • Takeno S, Noguchi T, Fumoto S, Kimura Y, Shibata T, Kawahara K. E-cadherin expression in patients with esophageal squamous cell carcinoma: promoter hypermethylation, Snail overexpression, and clinicopathologic implications. Am J Clin Pathol. 2004;122:78–84.

    Article  CAS  PubMed  Google Scholar 

  • Thiery JP. Epithelial–mesenchymal transitions in tumour progression. Nat Rev Cancer. 2002;2:442–54.

    Article  CAS  PubMed  Google Scholar 

  • Thiery JP, Acloque H, Huang RY, Nieto MA. Epithelial–mesenchymal transitions in development and disease. Cell. 2009;139:871–90.

    Article  CAS  PubMed  Google Scholar 

  • Torring N, Borre M, Sorensen KD, Andersen CL, Wiuf C, Orntoft TF. Genome-wide analysis of allelic imbalance in prostate cancer using the Affymetrix 50 K SNP mapping array. Br J Cancer. 2007;96:499–506.

    Article  CAS  PubMed  Google Scholar 

  • van der Neut R, Krimpenfort P, Calafat J, Niessen CM, Sonnenberg A. Epithelial detachment due to absence of hemidesmosomes in integrin beta 4 null mice. Nat Genet. 1996;13:366–9.

    Article  PubMed  Google Scholar 

  • Vega S, Morales AV, Ocana OH, Valdes F, Fabregat I, Nieto MA. Snail blocks the cell cycle and confers resistance to cell death. Genes Dev. 2004;18:1131–43.

    Article  CAS  PubMed  Google Scholar 

  • Waldmann J, Feldmann G, Slater EP, Langer P, Buchholz M, Ramaswamy A, et al. Expression of the zinc-finger transcription factor Snail in adrenocortical carcinoma is associated with decreased survival. Br J Cancer. 2008;99:1900–7.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • Yang MH, Chang SY, Chiou SH, Liu CJ, Chi CW, Chen PM, et al. Overexpression of NBS1 induces epithelial–mesenchymal transition and co-expression of NBS1 and Snail predicts metastasis of head and neck cancer. Oncogene. 2007;26:1459–67.

    Article  CAS  PubMed  Google Scholar 

  • Zutter MM, Santoro SA, Painter AS, Tsung YL, Gafford A. human alpha 2 integrin gene promoter. Identification of positive and negative regulatory elements important for cell-type and developmentally restricted gene expression. J Biol Chem. 1994;269:463–9.

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We thank Dr. Parvaneh Nikpour and Christiane Hader for their assistance in some experiments and helpful suggestions and Dr. Volker Jung, Homburg, for providing the PC-3 daughter cell lines. This research was supported by National Institute of Genetic Engineering and Biotechnology through the grant no. 218. We also would like to acknowledge and extend our gratitude to Iranian Blood Transfusion Organization, especially Dr. Ali Talebian and Dr. Ahmad Gharabaghian, for their contribution to this study.

Author information

Authors and Affiliations

  1. Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran

    Modjtaba Emadi Baygi

  2. Department of Biochemistry, National Institute of Genetic Engineering and Biotechnology, P.O. Box 14965/161, Tehran, Iran

    Zahra Soheila Soheili

  3. Institute for Molecular and Clinical Immunology, Otto-von-Guericke-University, Magdeburg, Germany

    Ingo Schmitz

  4. Helmholtz Center for Infection, Braunschweig, Germany

    Ingo Schmitz

  5. Iranian Blood Transfusion Research Center, Tehran, Iran

    Shahram Sameie

  6. Department of Urology, Heinrich Heine University, Düsseldorf, Germany

    Wolfgang A. Schulz

Authors
  1. Modjtaba Emadi Baygi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zahra Soheila Soheili
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ingo Schmitz
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shahram Sameie
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Wolfgang A. Schulz
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zahra Soheila Soheili.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Emadi Baygi, M., Soheili, Z.S., Schmitz, I. et al. Snail regulates cell survival and inhibits cellular senescence in human metastatic prostate cancer cell lines. Cell Biol Toxicol 26, 553–567 (2010). https://doi.org/10.1007/s10565-010-9163-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10565-010-9163-5

Keywords

Search

Navigation