Clinical & Experimental Metastasis

, Volume 28, Issue 5, pp 479–491 | Cite as

Impact of combined HDAC and mTOR inhibition on adhesion, migration and invasion of prostate cancer cells

  • Steffen Wedel
  • Lukasz Hudak
  • Jens-Michael Seibel
  • Jasmina Makarević
  • Eva Juengel
  • Igor Tsaur
  • Christoph Wiesner
  • Axel Haferkamp
  • Roman A. Blaheta
Research Paper


The concept of molecular tumor targeting might provide new hope in the treatment of advanced prostate cancer. We evaluated metastasis blocking properties of the histone deacetylase (HDAC) inhibitor valproic acid (VPA) and the mammalian target of rapamycin (mTOR) inhibitor RAD001 on prostate cancer cell lines. RAD001 or VPA were applied to PC-3 or LNCaP cells, either separately or in combination. Adhesion to vascular endothelium or to immobilized collagen, fibronectin or laminin was quantified. Migration and invasion were explored by a modified Boyden chamber assay. Integrin α and β subtypes were analyzed by flow cytometry, western blotting and RT-PCR. Effects of drug treatment on integrin related signaling, Akt and p70S6kinase activation, histone H3 and H4 acetylation were also determined. Separate application of RAD001 or VPA distinctly reduced tumor cell adhesion, migration and invasion, accompanied by elevated Akt activation and p70S6kinase de-activation. Integrin subtype expression was altered significantly by both compounds (VPA > RAD001). When both drugs were used in concert additive effects were observed on the migratory and invasive behavior but not on tumor-endothelium and tumor-matrix interaction. Separate mTOR or HDAC inhibition slows processes related to tumor metastasis. The RAD001-VPA combination showed advantage over VPA monotreatment with particular respect to migration and invasion. Ongoing studies are required to assess the relevance of VPA monotherapy versus VPA-RAD001 combination on tumor cell motility.


RAD001 Valproic acid Prostate carcinoma Invasion Adhesion 



Histone deacetylase


Valproic acid


Mammalian target of rapamycin


Human endothelial cells


Integrin-linked kinase


Focal adhesion kinase



We would like to thank Karen Nelson for critically reading the manuscript. This work was supported by the “Jung-Stiftung”.


  1. 1.
    Jemal A, Siegel R, Ward E et al (2008) Cancer statistics, 2008. CA Cancer J Clin 58:71–96PubMedCrossRefGoogle Scholar
  2. 2.
    Ward JF, Moul JW (2005) Rising prostate-specific antigen after primary prostate cancer therapy. Nat Clin Pract Urol 2:174–182PubMedCrossRefGoogle Scholar
  3. 3.
    Han M, Partin AW, Pound CR et al (2001) Long-term biochemical disease-free and cancer-specific survival following anatomic radical retropubic prostatectomy. The 15-year Johns Hopkins experience. Urol Clin North Am 28:555–565PubMedCrossRefGoogle Scholar
  4. 4.
    Wang L, Zou X, Berger AD et al (2009) Increased expression of histone deacetylaces (HDACs) and inhibition of prostate cancer growth and invasion by HDAC inhibitor SAHA. Am J Transl Res 1:62–71PubMedGoogle Scholar
  5. 5.
    Weichert W, Röske A, Gekeler V et al (2008) Histone deacetylases 1, 2 and 3 are highly expressed in prostate cancer and HDAC2 expression is associated with shorter PSA relapse time after radical prostatectomy. Br J Cancer 98:604–610PubMedCrossRefGoogle Scholar
  6. 6.
    Halkidou K, Gaughan L, Cook S et al (2004) Upregulation and nuclear recruitment of HDAC1 in hormone refractory prostate cancer. Prostate 59:177–189PubMedCrossRefGoogle Scholar
  7. 7.
    Kremer CL, Klein RR, Mendelson J et al (2006) Expression of mTOR signaling pathway markers in prostate cancer progression. Prostate 66:1203–1212PubMedCrossRefGoogle Scholar
  8. 8.
    Dai B, Kong YY, Ye DW et al (2009) Activation of the mammalian target of rapamycin signalling pathway in prostate cancer and its association with patient clinicopathological characteristics. BJU Int 104:1009–1016PubMedCrossRefGoogle Scholar
  9. 9.
    Bedolla R, Prihoda TJ, Kreisberg JI et al (2007) Determining risk of biochemical recurrence in prostate cancer by immunohistochemical detection of PTEN expression and Akt activation. Clin Cancer Res 13:3860–3867PubMedCrossRefGoogle Scholar
  10. 10.
    Shimizu Y, Segawa T, Inoue T et al (2007) Increased Akt and phosphorylated Akt expression are associated with malignant biological features of prostate cancer in Japanese men. BJU Int 100:685–690PubMedCrossRefGoogle Scholar
  11. 11.
    Angelucci A, Muzi P, Cristiano L et al (2008) Neuroendocrine transdifferentiation induced by VPA is mediated by PPARgamma activation and confers resistance to antiblastic therapy in prostate carcinoma. Prostate 68:588–598PubMedCrossRefGoogle Scholar
  12. 12.
    Shabbeer S, Kortenhorst MS, Kachhap S et al (2007) Multiple molecular pathways explain the anti-proliferative effect of valproic acid on prostate cancer cells in vitro and in vivo. Prostate 67:1099–1110PubMedCrossRefGoogle Scholar
  13. 13.
    Xia Q, Sung J, Chowdhury W et al (2006) Chronic administration of valproic acid inhibits prostate cancer cell growth in vitro and in vivo. Cancer Res 66:7237–7244PubMedCrossRefGoogle Scholar
  14. 14.
    Morgan TM, Pitts TE, Gross TS et al (2008) RAD001 (Everolimus) inhibits growth of prostate cancer in the bone and the inhibitory effects are increased by combination with docetaxel and zoledronic acid. Prostate 68:861–871PubMedCrossRefGoogle Scholar
  15. 15.
    Schayowitz A, Sabnis G, Njar VC et al (2008) Synergistic effect of a novel antiandrogen, VN/124-1, and signal transduction inhibitors in prostate cancer progression to hormone independence in vitro. Mol Cancer Ther 7:121–132PubMedCrossRefGoogle Scholar
  16. 16.
    Iiizumi M, Mohinta S, Bandyopadhyay S et al (2007) Tumor-endothelial cell interactions: therapeutic potential. Microvasc Res 74:114–120PubMedCrossRefGoogle Scholar
  17. 17.
    Hall CL, Dai J, van Golen KL et al (2006) Type I collagen receptor (alpha 2 beta 1) signaling promotes the growth of human prostate cancer cells within the bone. Cancer Res 66:8648–8654PubMedCrossRefGoogle Scholar
  18. 18.
    Van Slambrouck S, Jenkins AR, Romero AE et al (2009) Reorganization of the integrin alpha2 subunit controls cell adhesion and cancer cell invasion in prostate cancer. Int J Oncol 34:1717–1726PubMedGoogle Scholar
  19. 19.
    Liu JM, Bignon J, Haroun-Bouhedja F et al (2005) Inhibitory effect of fucoidan on the adhesion of adenocarcinoma cells to fibronectin. Anticancer Res 25:2129–2133PubMedGoogle Scholar
  20. 20.
    Chao WT, Kunz J (2009) Focal adhesion disassembly requires clathrin-dependent endocytosis of integrins. FEBS Lett 583:1337–1343Google Scholar
  21. 21.
    Oertl A, Relja B, Makarevic J et al (2006) Altered expression of beta1 integrins in renal carcinoma cell lines exposed to the differentiation inducer valproic acid. Int J Mol Med 18:347–354PubMedGoogle Scholar
  22. 22.
    Björkman M, Iljin K, Halonen P et al (2008) Defining the molecular action of HDAC inhibitors and synergism with androgen deprivation in ERG-positive prostate cancer. Int J Cancer 123:2774–2781PubMedCrossRefGoogle Scholar
  23. 23.
    Nagakawa O, Akashi T, Hayakawa Y et al (2004) Differential expression of integrin subunits in DU-145/AR prostate cancer cells. Oncol Rep 12:837–841PubMedGoogle Scholar
  24. 24.
    Welsbie DS, Xu J, Chen Y et al (2009) Histone deacetylases are required for androgen receptor function in hormone-sensitive and castrate-resistant prostate cancer. Cancer Res 69:958–966PubMedCrossRefGoogle Scholar
  25. 25.
    Slack-Davis JK, Parsons JT (2004) Emerging views of integrin signaling: implications for prostate cancer. J Cell Biochem 91:41–46PubMedCrossRefGoogle Scholar
  26. 26.
    Schmelz M, Cress AE, Scott KM et al (2002) Different phenotypes in human prostate cancer: alpha6 or alpha3 integrin in cell-extracellular adhesion sites. Neoplasia 4:243–254PubMedCrossRefGoogle Scholar
  27. 27.
    Lai TH, Fong YC, Fu WM et al (2008) Osteoblasts-derived BMP-2 enhances the motility of prostate cancer cells via activation of integrins. Prostate 68:1341–1353PubMedCrossRefGoogle Scholar
  28. 28.
    Wang X, Ferreira AM (2005) Shao Q et al. (2005) Beta3 integrins facilitate matrix interactions during transendothelial migration of PC3 prostate tumor cells. Prostate 63:65–80PubMedCrossRefGoogle Scholar
  29. 29.
    Balasubramanian S, Kuppuswamy D (2003) RGD-containing peptides activate S6K1 through beta3 integrin in adult cardiac muscle cells. J Biol Chem 278:42214–42224PubMedCrossRefGoogle Scholar
  30. 30.
    Priulla M, Calastretti A, Bruno P et al (2007) Preferential chemosensitization of PTEN-mutated prostate cells by silencing the Akt kinase. Prostate 67:782–789PubMedCrossRefGoogle Scholar
  31. 31.
    Verheul HM, Salumbides B, Van Erp K et al (2008) Combination strategy targeting the hypoxia inducible factor-1 alpha with mammalian target of rapamycin and histone deacetylase inhibitors. Clin Cancer Res 14:3589–3597PubMedCrossRefGoogle Scholar
  32. 32.
    Mahalingam D, Medina EC, Esquivel JA 2nd et al (2010) Vorinostat enhances the activity of temsirolimus in renal cell carcinoma through suppression of survivin levels. Clin Cancer Res 16:141–153PubMedCrossRefGoogle Scholar
  33. 33.
    Taliaferro-Smith L, Nagalingam A, Zhong D et al (2009) LKB1 is required for adiponectin-mediated modulation of AMPK-S6K axis and inhibition of migration and invasion of breast cancer cells. Oncogene 28:2621–2633PubMedCrossRefGoogle Scholar
  34. 34.
    Chen CS, Weng SC, Tseng PH et al (2005) Histone acetylation-independent effect of histone deacetylase inhibitors on Akt through the reshuffling of protein phosphatase 1 complexes. J Biol Chem 280:38879–38887PubMedCrossRefGoogle Scholar
  35. 35.
    Ridolfi E, Matteucci E, Maroni P et al (2008) Inhibitory effect of HGF on invasiveness of aggressive MDA-MB231 breast carcinoma cells, and role of HDACs. Br J Cancer 99:1623–1634PubMedCrossRefGoogle Scholar
  36. 36.
    Lydolph MC, Morgan-Fisher M, Høye AM et al (2009) Alpha9beta1 integrin in melanoma cells can signal different adhesion states for migration and anchorage. Exp Cell Res 315:3312–3324PubMedCrossRefGoogle Scholar
  37. 37.
    Lakshman M, Xu L, Ananthanarayanan V et al (2008) Dietary genistein inhibits metastasis of human prostate cancer in mice. Cancer Res 68:2024–2032PubMedCrossRefGoogle Scholar
  38. 38.
    Russell MR, Jamieson WL, Dolloff NG, Fatatis A (2009) The alpha-receptor for platelet-derived growth factor as a target for antibody-mediated inhibition of skeletal metastases from prostate cancer cells. Oncogene 28:412–421PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Steffen Wedel
    • 1
  • Lukasz Hudak
    • 1
  • Jens-Michael Seibel
    • 1
  • Jasmina Makarević
    • 1
  • Eva Juengel
    • 1
  • Igor Tsaur
    • 1
  • Christoph Wiesner
    • 1
  • Axel Haferkamp
    • 1
  • Roman A. Blaheta
    • 1
  1. 1.Department of UrologyJohann Wolfgang Goethe-UniversityFrankfurt am MainGermany

Personalised recommendations