Skip to main content

Advertisement

SpringerLink
Log in

The small conductance calcium-activated potassium channel 3 (SK3) is a molecular target for Edelfosine to reduce the invasive potential of urothelial carcinoma cells

  • Original Article
  • Published:
Tumor Biology

Abstract

Metastasis is the survival-determining factor in urothelial carcinoma (UC) of the urinary bladder. The small conductance calcium-activated potassium channel 3 (SK3) enhances tumor cell invasion in breast cancer and malignant melanoma. Since Edelfosine, a glycerophospholipid with antitumoral properties, effectively inhibits SK3 channel activity, our goal was to evaluate SK3 as a potential molecular target to inhibit the gain of an invasive phenotype in UC. SK3 protein expression was analyzed in 208 tissue samples and UC cell lines. Effects of Edelfosine on SK3 expression and intracellular calcium levels as well as on cell morphology, cell survival and proliferation were assessed using immunoblotting, potentiometric fluorescence microscopy, and clonogenic/cell survival assay; furthermore, we analyzed the effect of Edelfosine and SK3 RNAi knockdown on tumor cell migration and invasion in vitro and in vivo. We found that SK3 is strongly expressed in muscle-invasive UC and in the RT112 cellular tumor model. Higher concentrations of Edelfosine have a strong antitumoral effect on UC cells, while 1 μM effectively inhibits migration/invasion of UC cells in vitro and in vivo comparable to the SK3 knockdown phenotype. Taken together, our results show strong expression of SK3 in muscle-invasive UC, consistent with the postulated role of the protein in tumor cell invasion. Edelfosine is able to effectively inhibit migration and invasion of UC cells in vitro and in vivo in an SK3-dependent way, pointing towards a possible role for Edelfosine as an antiinvasive drug to effectively inhibit UC cell invasion and metastasis.

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

Similar content being viewed by others

References

  1. Burger M, Catto JW, Dalbagni G, Grossman HB, Herr H, Karakiewicz P, et al. Epidemiology and risk factors of urothelial bladder cancer. Eur Urol. 2013;63(2):234–41.

    Article  PubMed  Google Scholar 

  2. Bajorin DF, Dodd PM, Mazumdar M, Fazzari M, McCaffrey JA, Scher HI, et al. Long-term survival in metastatic transitional-cell carcinoma and prognostic factors predicting outcome of therapy. J Clin Oncol. 1999;17(10):3173–81.

    Article  CAS  PubMed  Google Scholar 

  3. Stenzl A, Cowan NC, De Santis M, Kuczyk MA, Merseburger AS, Ribal MJ, et al. Treatment of muscle-invasive and metastatic bladder cancer: update of the EAU guidelines. Eur Urol. 2011;59(6):1009–18.

    Article  CAS  PubMed  Google Scholar 

  4. Girault A, Haelters JP, Potier-Cartereau M, Chantome A, Jaffres PA, Bougnoux P, et al. Targeting SKCa channels in cancer: potential new therapeutic approaches. Curr Med Chem. 2012;19(5):697–713.

    Article  CAS  PubMed  Google Scholar 

  5. Liebau S, Steinestel J, Linta L, Kleger A, Storch A, Schoen M, et al. An SK3 channel/nWASP/Abi-1 complex is involved in early neurogenesis. PLoS One. 2011;6(3):e18148. doi:10.1371/journal.pone.0018148.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Liebau S, Vaida B, Proepper C, Grissmer S, Storch A, Boeckers TM, et al. Formation of cellular projections in neural progenitor cells depends on SK3 channel activity. J Neurochem. 2007;101(5):1338–50. doi:10.1111/j.1471-4159.2006.04437.x.

    Article  CAS  PubMed  Google Scholar 

  7. Bond CT, Maylie J, Adelman JP. SK channels in excitability, pacemaking and synaptic integration. Curr Opin Neurobiol. 2005;15(3):305–11.

    Article  CAS  PubMed  Google Scholar 

  8. Xia X-M, Fakler B, Rivard A, Wayman G, Johnson-Pais T, Keen J, et al. Mechanism of calcium gating in small-conductance calcium-activated potassium channels. Nature. 1998;395(6701):503–7.

    Article  CAS  PubMed  Google Scholar 

  9. Chantome A, Potier-Cartereau M, Clarysse L, Fromont G, Marionneau-Lambot S, Gueguinou M, et al. Pivotal role of the lipid Raft SK3-Orai1 complex in human cancer cell migration and bone metastases. Cancer Res. 2013;73(15):4852–61. doi:10.1158/0008-5472.can-12-4572.

    Article  CAS  PubMed  Google Scholar 

  10. Prevarskaya N, Skryma R, Shuba Y. Calcium in tumour metastasis: new roles for known actors. Nat Rev Cancer. 2011;11(8):609–18.

    Article  CAS  PubMed  Google Scholar 

  11. Steinestel K, Wardelmann E, Hartmann W, Grünewald I. Regulators of actin dynamics in gastrointestinal tract tumors. Gastroenterol Res Pract. 2015;2015:930157. doi:10.1155/2015/930157.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Potier M, Joulin V, Roger S, Besson P, Jourdan ML, Leguennec JY, et al. Identification of SK3 channel as a new mediator of breast cancer cell migration. Mol Cancer Ther. 2006;5(11):2946–53. doi:10.1158/1535-7163.MCT-06-0194.

    Article  CAS  PubMed  Google Scholar 

  13. Chantome A, Girault A, Potier M, Collin C, Vaudin P, Pagès J-C, et al. KCa2.3 channel-dependent hyperpolarization increases melanoma cell motility. Exp Cell Res. 2009;315(20):3620–30. doi:10.1016/j.yexcr.2009.07.021.

    Article  CAS  PubMed  Google Scholar 

  14. Potier M, Chantôme A, Joulin V, Girault A, Roger S, Besson P, et al. The SK3/KCa2.3 potassium channel is a new cellular target for edelfosine. Br J Pharmacol. 2011;162(2):464–79.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Slaton JW, Hampton JA, Selman SH. Exposure to alkyllysophospholipids inhibits in vitro invasion of transitional cell carcinoma. J Urol. 1994;152(5 Pt 1):1594–8.

    CAS  PubMed  Google Scholar 

  16. Steinestel J, Cronauer M, Müller J, Al Ghazal A, Skowronek P, Arndt A, et al. Overexpression of p16 (INK4a) in urothelial carcinoma in situ is a marker for MAPK-mediated epithelial-mesenchymal transition but is not related to human papillomavirus infection. PLoS One. 2013;8(5):e65189-e.

    Article  Google Scholar 

  17. Innocenti M, Gerboth S, Rottner K, Lai FP, Hertzog M, Stradal TE, et al. Abi1 regulates the activity of N-WASP and WAVE in distinct actin-based processes. Nat Cell Biol. 2005;7(10):969–76.

    Article  CAS  PubMed  Google Scholar 

  18. Steinestel K, Brüderlein S, Lennerz JK, Steinestel J, Kraft K, Pröpper C, et al. Expression and Y435-phosphorylation of Abelson interactor 1 (Abi1) promotes tumour cell adhesion, extracellular matrix degradation and invasion by colorectal carcinoma cells. Mol Cancer. 2014;13(1):145.

    Article  PubMed  PubMed Central  Google Scholar 

  19. Weaver AK, Bomben VC, Sontheimer H. Expression and function of calcium‐activated potassium channels in human glioma cells. Glia. 2006;54(3):223–33.

    Article  PubMed  PubMed Central  Google Scholar 

  20. Witjes JA, Compérat E, Cowan NC, De Santis M, Gakis G, Lebret T, et al. EAU guidelines on muscle-invasive and metastatic bladder cancer: summary of the 2013 guidelines. Eur Urol. 2014;65(4):778–92.

    Article  PubMed  Google Scholar 

  21. Haller T, Dietl P, Deetjen P, Völkl H. The lysosomal compartment as intracellular calcium store in MDCK cells: a possible involvement in InsP 3-mediated Ca 2+ release. Cell Calcium. 1996;19(2):157–65.

    Article  CAS  PubMed  Google Scholar 

  22. Isachenko V, Mallmann P, Petrunkina AM, Rahimi G, Nawroth F, Hancke K, et al. Comparison of in vitro-and chorioallantoic membrane (CAM)-culture systems for cryopreserved medulla-contained human ovarian tissue. PLoS One. 2012;7(3):e32549.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

The authors would like to express their gratitude to Professor Axel Zander, Cellprotect & Co. KG (Eutin, Germany), for the kind gift of Edelfosine in pharmaceutical purity.

The work of KS is supported by the Deutsche Forschungsgemeinschaft (grant STE 2467/1-1) and by the Medical Faculty of the University of Münster (IMF grant I-SP111504).

Author information

Author notes

    Authors and Affiliations

    1. Gerhard Domagk Institute of Pathology, University of Münster, Domagkstrasse 17, 48149, Münster, Germany

      Konrad Steinestel & Eva Wardelmann

    2. Bundeswehr Institute of Radiobiology, Neuherbergstr. 11, 80937, Munich, Germany

      Stefan Eder

    3. Department of General Physiology, University of Ulm, Albert-Einstein-Allee 11, 89081, Ulm, Germany

      Konstantin Ehinger

    4. Institute of Pathology and Molecular Pathology, Bundeswehrkrankenhaus Ulm, Oberer Eselsberg 40, 89081, Ulm, Germany

      Juliane Schneider

    5. Department of Urology, Ulm University Medical Center, Prittwitzstrasse 43, 89075, Ulm, Germany

      Felicitas Genze & Eva Winkler

    6. Clinic of Urology, Münster University Medical Center, Albert-Schweitzer-Campus 1, Building A1, 48149, Münster, Germany

      Andres J. Schrader & Julie Steinestel

    Authors
    1. Konrad Steinestel
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Stefan Eder
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Konstantin Ehinger
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Juliane Schneider
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Felicitas Genze
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Eva Winkler
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. Eva Wardelmann
      View author publications

      You can also search for this author in PubMed Google Scholar

    8. Andres J. Schrader
      View author publications

      You can also search for this author in PubMed Google Scholar

    9. Julie Steinestel
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Konrad Steinestel.

    Ethics declarations

    Conflict of interest

    None

    Additional information

    Konrad Steinestel and Stefan Eder contributed equally to this work.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Steinestel, K., Eder, S., Ehinger, K. et al. The small conductance calcium-activated potassium channel 3 (SK3) is a molecular target for Edelfosine to reduce the invasive potential of urothelial carcinoma cells. Tumor Biol. 37, 6275–6283 (2016). https://doi.org/10.1007/s13277-015-4509-5

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s13277-015-4509-5

    Keywords

    Search

    Navigation