Tumor Biology

, Volume 36, Issue 6, pp 4861–4869 | Cite as

Effects of zoledronic acid and docetaxel on small GTP-binding proteins in prostate cancer

  • Rachel M. Jones
  • Claire Morgan
  • Gianfilippo Bertelli
Research Article


Increasingly, in castration-resistant prostate cancer, patients are often treated with docetaxel and the bisphosphonate zoledronic acid concurrently, yet there is still a paucity in the literature regarding the molecular basis of how this drug combination works. The study was performed on the hormone-resistant cell line PC-3. Cells were treated with clinically relevant concentrations of docetaxel and zoledronic acid either as single agents or in sequence and combination. Cell viability and apoptosis were assessed along with the prenylation status of the GTPases Ras and RhoA. Following 1-mM zoledronic acid treatment, inhibition of the prenylation of H-Ras and Rho A was observed along with an increase in the unprenylated form in the cytoplasm. Docetaxel 1 nM and zoledronic acid 1 mM also showed an increase in the unprenylated form of both small GTP-binding proteins in the cytoplasm and a reduction of protein in the membrane fraction. Overall, zoledronic acid followed by docetaxel was the best regimen producing the greatest reduction in cell viability and increase in apoptosis. At the highest concentrations of zoledronic acid and docetaxel, zoledronic acid followed by docetaxel was also the most effective at reducing the prenylation of both H-Ras and RhoA at the membrane. We have demonstrated that clinically achievable concentrations of zoledronic acid and docetaxel cause a reduction in the prenylation of both H-Ras and Rho A and a reduction of protein movement into the membrane. The most effective regimen overall was high-dose zoledronic acid followed by docetaxel, suggesting that this regimen may be of benefit in clinical practice.


Castration resistant Prostate cancer RhoA Ras Zoledronic acid Docetaxel 


Conflicts of interest



  1. 1.
    Oades GM, Senaratne SG, Clarke IA, Kirby RS, Colston KW. Nitrogen containing bisphosphonates induce apoptosis and inhibit the mevalonate pathway, impairing Ras membrane localization in prostate cancer cells. J Urol. 2003;170(1):246–52.CrossRefPubMedGoogle Scholar
  2. 2.
    Han B, Fujimoto N, Kobayashi M, Matsumoto T. Synergistic effect of geranylgeranyltransferase inhibitor, GGTI, and docetaxel on the growth of prostate cancer cells. Prostate Cancer. 2012;2012:989214.CrossRefPubMedGoogle Scholar
  3. 3.
    Morgan C, Lewis PD, Jones RM, Bertelli G, Thomas GA, Leonard RC. The in vitro anti-tumour activity of zoledronic acid and docetaxel at clinically achievable concentrations in prostate cancer. Acta Oncol. 2007;46(5):669–77.CrossRefPubMedGoogle Scholar
  4. 4.
    Luckman SP, Hughes DE, Coxon FP, Graham R, Russell G, Rogers MJ. Nitrogen-containing bisphosphonates inhibit the mevalonate pathway and prevent post-translational prenylation of GTP-binding proteins, including Ras. J Bone Miner Res. 1998;13(4):581–9.CrossRefPubMedGoogle Scholar
  5. 5.
    van Beek E, Pieterman E, Cohen L, Lowik C, Papapoulos S. Farnesyl pyrophosphate synthase is the molecular target of nitrogen-containing bisphosphonates. Biochem Biophys Res Commun. 1999;264(1):108–11.CrossRefPubMedGoogle Scholar
  6. 6.
    Senaratne SG, Mansi JL, Colston KW. The bisphosphonate zoledronic acid impairs Ras membrane [correction of impairs membrane] localisation and induces cytochrome c release in breast cancer cells. Br J Cancer. 2002;86(9):1479–86.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Goffinet M, Thoulouzan M, Pradines A, Lajoie-Mazenc I, Weinbaum C, Faye JC, et al. Zoledronic acid treatment impairs protein geranyl-geranylation for biological effects in prostatic cells. BMC Cancer. 2006;6:60.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    McCubrey JA, Steelman LS, Chappell WH, Abrams SL, Wong EW, Chang F, et al. Roles of the Raf/MEK/ERK pathway in cell growth, malignant transformation and drug resistance. Biochim Biophys Acta. 2007;1773(8):1263–84.CrossRefPubMedGoogle Scholar
  9. 9.
    Kolch W. Meaningful relationships: the regulation of the Ras/Raf/MEK/ERK pathway by protein interactions. Biochem J. 2000;351(Pt 2):289–305.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Coxon JP, Oades GM, Kirby RS, Colston KW. Zoledronic acid induces apoptosis and inhibits adhesion to mineralized matrix in prostate cancer cells via inhibition of protein prenylation. BJU Int. 2004;94(1):164–70.CrossRefPubMedGoogle Scholar
  11. 11.
    Nogawa M, Yuasa T, Kimura S, Kuroda J, Segawa H, Sato K, et al. Zoledronic acid mediates Ras-independent growth inhibition of prostate cancer cells. Oncol Res. 2005;15(1):1–9.PubMedGoogle Scholar
  12. 12.
    Jagdev SP, Coleman RE, Shipman CM, Rostami HA, Croucher PI. The bisphosphonate, zoledronic acid, induces apoptosis of breast cancer cells: evidence for synergy with paclitaxel. Br J Cancer. 2001;84(8):1126–34.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Ringel I, Horwitz SB. Studies with RP 56976 (taxotere): a semisynthetic analogue of taxol. J Natl Cancer Inst. 1991;83(4):288–91.CrossRefPubMedGoogle Scholar
  14. 14.
    Ito T, Deng X, Carr B, May WS. Bcl-2 phosphorylation required for anti-apoptosis function. J Biol Chem. 1997;272(18):11671–3.CrossRefPubMedGoogle Scholar
  15. 15.
    Sweeney CJ, Miller KD, Sissons SE, Nozaki S, Heilman DK, Shen J, et al. The antiangiogenic property of docetaxel is synergistic with a recombinant humanized monoclonal antibody against vascular endothelial growth factor or 2-methoxyestradiol but antagonized by endothelial growth factors. Cancer Res. 2001;61(8):3369–72.PubMedGoogle Scholar
  16. 16.
    Danesi R, Figg WD, Reed E, Myers CE. Paclitaxel (taxol) inhibits protein isoprenylation and induces apoptosis in PC-3 human prostate cancer cells. Mol Pharmacol. 1995;47(6):1106–11.PubMedGoogle Scholar
  17. 17.
    Moasser MM, Sepp-Lorenzino L, Kohl NE, Oliff A, Balog A, Su DS, et al. Farnesyl transferase inhibitors cause enhanced mitotic sensitivity to taxol and epothilones. Proc Natl Acad Sci U S A. 1998;95(4):1369–74.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Appels NM, Beijnen JH, Schellens JH. Development of farnesyl transferase inhibitors: a review. Oncologist. 2005;10(8):565–78.CrossRefPubMedGoogle Scholar
  19. 19.
    Yamamoto N, Tamura T, Murakami H, Shimoyama T, Nokihara H, Ueda Y, et al. Randomized pharmacokinetic and pharmacodynamic study of docetaxel: dosing based on body-surface area compared with individualized dosing based on cytochrome P450 activity estimated using a urinary metabolite of exogenous cortisol. J Clin Oncol. 2005;23(6):1061–9.CrossRefPubMedGoogle Scholar
  20. 20.
    Holstein SA, Wohlford-Lenane CL, Hohl RJ. Consequences of mevalonate depletion. Differential transcriptional, translational, and post-translational up-regulation of Ras, Rap1a, RhoA, AND RhoB. J Biol Chem. 2002;277(12):10678–82.CrossRefPubMedGoogle Scholar
  21. 21.
    Lerner EC, Qian Y, Blaskovich MA, Fossum RD, Vogt A, Sun J, et al. Ras CAAX peptidomimetic FTI-277 selectively blocks oncogenic Ras signaling by inducing cytoplasmic accumulation of inactive Ras-Raf complexes. J Biol Chem. 1995;270(45):26802–6.CrossRefPubMedGoogle Scholar
  22. 22.
    Reyes-Moreno C, Sourla A, Choki I, Doillon C, Koutsilieris M. Osteoblast-derived survival factors protect PC-3 human prostate cancer cells from adriamycin apoptosis. Urology. 1998;52(2):341–7.CrossRefPubMedGoogle Scholar
  23. 23.
    Li X, Liu L, Tupper JC, Bannerman DD, Winn RK, Sebti SM, et al. Inhibition of protein geranylgeranylation and RhoA/RhoA kinase pathway induces apoptosis in human endothelial cells. J Biol Chem. 2002;277(18):15309–16.CrossRefPubMedGoogle Scholar
  24. 24.
    Denoyelle C, Hong L, Vannier JP, Soria J, Soria C. New insights into the actions of bisphosphonate zoledronic acid in breast cancer cells by dual RhoA-dependent and -independent effects. Br J Cancer. 2003;88(10):1631–40.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Sato M, Grasser W, Endo N, Akins R, Simmons H, Thompson DD, et al. Bisphosphonate action. Alendronate localization in rat bone and effects on osteoclast ultrastructure. J Clin Invest. 1991;88(6):2095–105.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Gioeli D. Signal transduction in prostate cancer progression. Clin Sci (Lond). 2005;108(4):293–308.CrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Rachel M. Jones
    • 1
  • Claire Morgan
    • 2
  • Gianfilippo Bertelli
    • 3
  1. 1.Gynaecology DepartmentVelindre Cancer CentreCardiffUK
  2. 2.College of MedicineSwansea UniversitySwanseaUK
  3. 3.South West Wales Cancer InstituteSingleton HospitalSwanseaUK

Personalised recommendations