Tumor Biology

, Volume 37, Issue 1, pp 1025–1033 | Cite as

Preclinical evaluation of perifosine as a potential promising anti-rhabdomyosarcoma agent

Research Article


Rhabdomyosarcoma (RMS) is a highly malignant and metastatic pediatric cancer that arises from the skeletal muscle. Recent studies have identified an important role of AKT signaling in RMS progression. In the current study, we investigated the activity of perifosine, an oral alkylphospholipid AKT inhibitor, against human RMS cells (RD and Rh-30 lines) both in vivo and in vitro, and studied the underlying mechanisms. We showed that perifosine significantly inhibited RMS cell growth in concentration- and time-dependent manners. Meanwhile, perifosine induced dramatic apoptosis in RMS cells. At the signaling level, perifosine blocked AKT activation, while inducing reactive oxygen species (ROS) production as well as JNK and P38 phosphorylations in RMS cells. Restoring AKT activation by introducing a constitutively active-AKT (CA-AKT) only alleviated (not abolished) perifosine-induced cytotoxicity in RD cells. Yet, the ROS scavenger N-acetyl cysteine (NAC) as well as pharmacological inhibitors against JNK (SP-600125) or P38 (SB-203580) suppressed perifosine-induced cytotoxicity in RMS cells. Thus, perifosine induces growth inhibition and apoptosis in RMS cells through mechanisms more than just blocking AKT. In vivo, oral administration of perifosine significantly inhibited growth of Rh-30 xenografts in severe combined immunodeficient (SCID) mice. Our data indicate that perifosine might be further investigated as a promising anti-RMS agent.


Rhabdomyosarcoma (RMS) AKT Perifosine Signaling and apoptosis 



This work is supported by the Project of Science and Technology Department of Zhejiang Province (2014C33206).

Conflicts of interest



  1. 1.
    Dagher R, Helman L. Rhabdomyosarcoma: an overview. Oncologist. 1999;4:34–44.PubMedGoogle Scholar
  2. 2.
    Merlino G, Helman LJ. Rhabdomyosarcoma—working out the pathways. Oncogene. 1999;18:5340–8.CrossRefPubMedGoogle Scholar
  3. 3.
    Spunt SL, Smith LM, Ruymann FB, Qualman SJ, Donaldson SS, Rodeberg DA, et al. Cyclophosphamide dose intensification during induction therapy for intermediate-risk pediatric rhabdomyosarcoma is feasible but does not improve outcome: a report from the soft tissue sarcoma committee of the children’s oncology group. Clin Cancer Res. 2004;10:6072–9.CrossRefPubMedGoogle Scholar
  4. 4.
    Crist WM, Anderson JR, Meza JL, Fryer C, Raney RB, Ruymann FB, et al. Intergroup rhabdomyosarcoma study-iv: Results for patients with nonmetastatic disease. J Clin Oncol. 2001;19:3091–102.CrossRefPubMedGoogle Scholar
  5. 5.
    Vanhaesebroeck B, Stephens L, Hawkins P. Pi3k signalling: the path to discovery and understanding. Nat Rev Mol Cell Biol. 2012;13:195–203.CrossRefPubMedGoogle Scholar
  6. 6.
    Fruman DA, Rommel C. Pi3k and cancer: lessons, challenges and opportunities. Nat Rev Drug Discov. 2014;13:140–56.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Petricoin 3rd EF, Espina V, Araujo RP, Midura B, Yeung C, Wan X, et al. Phosphoprotein pathway mapping: akt/mammalian target of rapamycin activation is negatively associated with childhood rhabdomyosarcoma survival. Cancer Res. 2007;67:3431–40.CrossRefPubMedGoogle Scholar
  8. 8.
    Cao L, Yu Y, Darko I, Currier D, Mayeenuddin LH, Wan X, et al. Addiction to elevated insulin-like growth factor i receptor and initial modulation of the akt pathway define the responsiveness of rhabdomyosarcoma to the targeting antibody. Cancer Res. 2008;68:8039–48.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Cirstea D, Hideshima T, Rodig S, Santo L, Pozzi S, Vallet S, et al. Dual inhibition of akt/mammalian target of rapamycin pathway by nanoparticle albumin-bound-rapamycin and perifosine induces antitumor activity in multiple myeloma. Mol Cancer Ther. 2010;9:963–75.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Kondapaka SB, Singh SS, Dasmahapatra GP, Sausville EA, Roy KK. Perifosine, a novel alkylphospholipid, inhibits protein kinase b activation. Mol Cancer Ther. 2003;2:1093–103.PubMedGoogle Scholar
  11. 11.
    Ruiter GA, Zerp SF, Bartelink H, van Blitterswijk WJ, Verheij M. Alkyl-lysophospholipids activate the sapk/jnk pathway and enhance radiation-induced apoptosis. Cancer Res. 1999;59:2457–63.PubMedGoogle Scholar
  12. 12.
    Fei HR, Chen G, Wang JM, Wang FZ. Perifosine induces cell cycle arrest and apoptosis in human hepatocellular carcinoma cell lines by blockade of akt phosphorylation. Cytotechnology. 2010;62:449–60.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Papa V, Tazzari PL, Chiarini F, Cappellini A, Ricci F, Billi AM, et al. Proapoptotic activity and chemosensitizing effect of the novel akt inhibitor perifosine in acute myelogenous leukemia cells. Leukemia. 2008;22:147–60.CrossRefPubMedGoogle Scholar
  14. 14.
    Crul M, Rosing H, de Klerk GJ, Dubbelman R, Traiser M, Reichert S, et al. Phase i and pharmacological study of daily oral administration of perifosine (d-21266) in patients with advanced solid tumours. Eur J Cancer. 2002;38:1615–21.CrossRefPubMedGoogle Scholar
  15. 15.
    Van Ummersen L, Binger K, Volkman J, Marnocha R, Tutsch K, Kolesar J, et al. A phase i trial of perifosine (nsc 639966) on a loading dose/maintenance dose schedule in patients with advanced cancer. Clin Cancer Res. 2004;10:7450–6.CrossRefPubMedGoogle Scholar
  16. 16.
    Zhang JL, Xu Y, Shen J. Cordycepin inhibits lipopolysaccharide (lps)-induced tumor necrosis factor (tnf)-alpha production via activating amp-activated protein kinase (ampk) signaling. Int J Mol Sci. 2014;15:12119–34.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Shen J, Liang L, Wang C. Perifosine inhibits lipopolysaccharide (lps)-induced tumor necrosis factor (tnf)-alpha production via regulation multiple signaling pathways: new implication for Kawasaki disease (kd) treatment. Biochem Biophys Res Commun. 2013;437:250–5.CrossRefPubMedGoogle Scholar
  18. 18.
    Sun H, Yu T, Li J. Co-administration of perifosine with paclitaxel synergistically induces apoptosis in ovarian cancer cells: more than just akt inhibition. Cancer Lett. 2011;310:118–28.CrossRefPubMedGoogle Scholar
  19. 19.
    Gills JJ, Dennis PA. Perifosine: update on a novel akt inhibitor. Curr Oncol Rep. 2009;11:102–10.CrossRefPubMedGoogle Scholar
  20. 20.
    Fensterle J, Aicher B, Seipelt I, Teifel M, Engel J. Current view on the mechanism of action of perifosine in cancer. Anti Cancer Agents Med Chem. 2014;14:629–35.CrossRefGoogle Scholar
  21. 21.
    Ji C, Yang YL, Yang Z, Tu Y, Cheng L, Chen B, et al. Perifosine sensitizes uvb-induced apoptosis in skin cells: new implication of skin cancer prevention? Cell Signal. 2012;24:1781–9.CrossRefPubMedGoogle Scholar
  22. 22.
    Chen MB, Wu XY, Tao GQ, Liu CY, Chen J, Wang LQ, et al. Perifosine sensitizes curcumin induced anti-colorectal cancer effects by targeting multiple signaling pathways both in vivo and in vitro. Int J Cancer. 2012;131(11):2487–98.CrossRefPubMedGoogle Scholar
  23. 23.
    Krawczyk J, Keane N, Swords R, O'Dwyer M, Freeman CL, Giles FJ. Perifosine—a new option in treatment of acute myeloid leukemia? Expert Opin Investig Drugs. 2013;22:1315–27.CrossRefPubMedGoogle Scholar
  24. 24.
    Srivastava N, Cho DC. Perifosine in renal cell carcinoma. Expert Opin Investig Drugs. 2013;22:285–91.CrossRefPubMedGoogle Scholar
  25. 25.
    Richardson PG, Eng C, Kolesar J, Hideshima T, Anderson KC. Perifosine, an oral, anti-cancer agent and inhibitor of the akt pathway: mechanistic actions, pharmacodynamics, pharmacokinetics, and clinical activity. Expert Opin Drug Metab Toxicol. 2012;8:623–33.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Vivanco I, Sawyers CL. The phosphatidylinositol 3-kinase akt pathway in human cancer. Nat Rev Cancer. 2002;2:489–501.CrossRefPubMedGoogle Scholar
  27. 27.
    Wang Q, Wang X, Hernandez A, Hellmich MR, Gatalica Z, Evers BM. Regulation of trail expression by the phosphatidylinositol 3-kinase/akt/gsk-3 pathway in human colon cancer cells. J Biol Chem. 2002;277:36602–10.CrossRefPubMedGoogle Scholar
  28. 28.
    Yao C, Wei JJ, Wang ZY, Ding HM, Li D, Yan SC, et al. Perifosine induces cell apoptosis in human osteosarcoma cells: new implication for osteosarcoma therapy? Cell Biochem Biophys. 2013;65:217–27.CrossRefPubMedGoogle Scholar
  29. 29.
    Tazzari PL, Tabellini G, Ricci F, Papa V, Bortul R, Chiarini F, et al. Synergistic proapoptotic activity of recombinant trail plus the akt inhibitor perifosine in acute myelogenous leukemia cells. Cancer Res. 2008;68:9394–403.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Rahmani M, Reese E, Dai Y, Bauer C, Payne SG, Dent P, et al. Coadministration of histone deacetylase inhibitors and perifosine synergistically induces apoptosis in human leukemia cells through akt and erk1/2 inactivation and the generation of ceramide and reactive oxygen species. Cancer Res. 2005;65:2422–32.CrossRefPubMedGoogle Scholar
  31. 31.
    Hideshima T, Catley L, Yasui H, Ishitsuka K, Raje N, Mitsiades C, et al. Perifosine, an oral bioactive novel alkylphospholipid, inhibits akt and induces in vitro and in vivo cytotoxicity in human multiple myeloma cells. Blood. 2006;107:4053–62.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Momota H, Nerio E, Holland EC. Perifosine inhibits multiple signaling pathways in glial progenitors and cooperates with temozolomide to arrest cell proliferation in gliomas in vivo. Cancer Res. 2005;65:7429–35.CrossRefPubMedGoogle Scholar
  33. 33.
    Ghobrial IM, Roccaro A, Hong F, Weller E, Rubin N, Leduc R, et al. Clinical and translational studies of a phase ii trial of the novel oral akt inhibitor perifosine in relapsed or relapsed/refractory Waldenstrom’s macroglobulinemia. Clin Cancer Res. 2010;16:1033–41.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Vink SR, Schellens JH, Beijnen JH, Sindermann H, Engel J, Dubbelman R, et al. Phase i and pharmacokinetic study of combined treatment with perifosine and radiation in patients with advanced solid tumours. Radiother Oncol. 2006;80:207–13.CrossRefPubMedGoogle Scholar
  35. 35.
    Knowling M, Blackstein M, Tozer R, Bramwell V, Dancey J, Dore N, et al. A phase ii study of perifosine (d-21226) in patients with previously untreated metastatic or locally advanced soft tissue sarcoma: a National Cancer Institute Of Canada Clinical Trials Group trial. Investig New Drugs. 2006;24:435–9.CrossRefGoogle Scholar
  36. 36.
    Pal I, Mandal M. Pi3k and akt as molecular targets for cancer therapy: current clinical outcomes. Acta Pharmacol Sin. 2012;33:1441–58.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  1. 1.Department of Respiratory DiseasesThe First Affiliated Hospital of Zhejiang UniversityHangzhouPeople’s Republic of China
  2. 2.Department of Thoracic OncologyThe First Affiliated Hospital of Zhejiang UniversityHangzhouPeople’s Republic of China

Personalised recommendations