Cancer Chemotherapy and Pharmacology

, Volume 62, Issue 2, pp 305–313 | Cite as

Antiproliferative effects of rapamycin as a single agent and in combination with carboplatin and paclitaxel in head and neck cancer cell lines

  • Nasredine Aissat
  • Christophe Le Tourneau
  • Aïda Ghoul
  • Maria Serova
  • Ivan Bieche
  • François Lokiec
  • Eric Raymond
  • Sandrine Faivre
Original Article



Recent data suggested that combining targeted therapies with chemotherapy may counteract drug resistance. Activation of the PI3K/AKT/mTOR pathway downstream to kinase receptors, such as EGFR, was found in 57–81% of head and neck squamous cell carcinoma (HNSCC), and was eventually associated with a loss of PTEN function. mTOR was shown to modulate cell proliferation, apoptosis, invasion, and angiogenesis. This study aimed to evaluate molecular and cellular effects of rapamycin in a panel of cell lines either as single agent or in combination with cytotoxics commonly used in HNSCC.


Antiproliferative effects of rapamycin, carboplatin, and paclitaxel were evaluated in a panel of three HNSCC cell lines (SCC61, SQ20B and HEP2). Cells were exposed to rapamycin for 48 h, to carboplatin for 48 h, or to paclitaxel for 24 h. Antiproliferative effects of simultaneous and sequential rapamycin-based combinations were studied using MTT assay and median effect plot analysis. Cell cycle effects were analysed using flow cytometry.


Rapamycin induced concentration dependent antiproliferative effects in HNSCC cell lines with IC50 of 5 ± 1, 12 ± 2 and 20 ± 2 μM in SCC61, SQ20B, and HEP2 cells, respectively. Higher antiproliferative effects were observed in SCC61 cells overexpressing NOXA and cyclin D1 than in HEP2 that overexpressed MDR1 and BCL2. In our panel, antiproliferative effects of rapamycin were associated with G0/G1 cell cycle accumulation and apoptosis induction, at concentrations ranging 3–30 μM. Combinations of rapamycin with paclitaxel and carboplatin displayed synergistic and additive effects. Synergistic effects were observed with paclitaxel in SQ20B and HEP2 cells and with carboplatin in SQ20B cells, when cells were exposed to cytotoxics prior to rapamycin.


Our results show that rapamycin displays antiproliferative effects and induces apoptosis in HNSCC cell lines, cellular effects being more potent in cells that do not express BCL2 and MDR1. Additive and synergistic effects were observed when rapamycin was combined with carboplatin and paclitaxel.


mTOR PI3K/AKT pathway Cytotoxicity Apoptosis 


  1. 1.
    Grandis JR, Tweardy DJ (1993) Elevated levels of transforming growth factor alpha and epidermal growth factor receptor messenger RNA are early markers of carcinogenesis in head and neck cancer. Cancer Res 53:3579–3584PubMedGoogle Scholar
  2. 2.
    Grandis JR, Melhem MF, Gooding WE et al (1998) Levels of TGF-alpha and EGFR protein in head and neck squamous cell carcinoma and patient survival. J Natl Cancer Inst 90:824–832CrossRefGoogle Scholar
  3. 3.
    Vivanco I, Sawyers CL (2002) The phosphatidylinositol 3-kinase AKT pathway in human cancer. Nat Rev Cancer 2:489–501PubMedCrossRefGoogle Scholar
  4. 4.
    Brown EJ, Albers MW, Shin TB et al (1994) A mammalian protein targeted by G1-arrestingreceptor complex. Nature 369:756–768PubMedCrossRefGoogle Scholar
  5. 5.
    Bjornsti MA, Houghton PJ (2004) The TOR pathway: a target for cancer therapy. Nat Rev Cancer 4:335–348PubMedCrossRefGoogle Scholar
  6. 6.
    Vignot S, Faivre S, Aguirre D et al (2005) mTOR-targeted therapy of cancer with rapamycin derivatives. Ann Oncol 16:525–537PubMedCrossRefGoogle Scholar
  7. 7.
    Bjornsti MA, Houghton PJ (2004) Lost in translation: dysregulation of cap-dependent translation and cancer. Cancer Cell 5:519–523PubMedCrossRefGoogle Scholar
  8. 8.
    Gera JF, Mellinghoff IK, Shi Y et al (2004) AKT activity determines sensitivity to mammalian target of rapamycin (mTOR) inhibitors by regulating cyclin D1 and c-myc expression. J Biol Chem 4:2737–2746Google Scholar
  9. 9.
    Faivre S, Kroemer G, Raymond E (2006) Current development of mTOR inhibitors as anticancer agents. Nat Rev Drug Discov 5(8):671–688PubMedCrossRefGoogle Scholar
  10. 10.
    Grewe M, Gansauge F, Schmid RM et al (1999) Regulation of cell growth and cyclin D1 expression by the constitutively active FRAP-p70s6K pathway in human pancreatic cancer cells. Cancer Res 59:3581–3587PubMedGoogle Scholar
  11. 11.
    Dong J, Peng J, Zhang H et al (2005) Role of glycogen synthase kinase 3beta in rapamycin-mediated cell cycle regulation and chemosensitivity. Cancer Res 65(5):1961–1972PubMedCrossRefGoogle Scholar
  12. 12.
    Oki E, Baba H, Tokunaga E et al (2005) Akt phosphorylation associates with LOH of PTEN and leads to chemoresistance for gastric cancer. Int J Cancer 117(3):376–380PubMedCrossRefGoogle Scholar
  13. 13.
    Geoerger B, Kerr K, Tang CB et al (2001) Antitumor activity of the Rapamycin analog CCI-779 in human primitive neuroectodermal Tumor/Medulloblastoma models as single agent and in combination Chemotherapy. Cancer Res 61:1527–1532PubMedGoogle Scholar
  14. 14.
    Gupta A, Dai Y, Vethanayagam RR et al (2006) Cyclosporin A, tacrolimus and sirolimus are potent inhibitors of the human breast cancer resistance protein (ABCG2) and reverse resistance to mitoxantrone and topotecan. Cancer Chemother Pharmacol 58(3):374–383PubMedCrossRefGoogle Scholar
  15. 15.
    Nagasawa I, Keng P, Maki C et al (1988) Absence of a radiation-induced first-cycle G1-S arrest in p53+ human tumor cells synchronized by mitotic selection. Cancer Res 58:2036–2041Google Scholar
  16. 16.
    Maggiorella L, Frascogna V, Poullain MG et al (2001) The Olivacine S16020 enhances the Antitumor effect of ionizing radiation without increasing radio-induced Mucositis. Clin Cancer Res 7:2091–2095PubMedGoogle Scholar
  17. 17.
    Brachman DG, Beckett M, Graves D et al (1993) p53 mutation does not correlate with radiosensitivity in 24 head and neck cancer cell lines. Cancer Res 53:3667–3669PubMedGoogle Scholar
  18. 18.
    Bièche I, Parfait B, Tozlu S et al (2001) Quantitation of androgen receptor gene expression in sporadic breast tumors by real-time RT-PCR: evidence that MYC is an AR-regulated gene. Carcinogenesis 22:1521–1526PubMedCrossRefGoogle Scholar
  19. 19.
    Chou TC, Talalay P (1984) Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul 22:27–55PubMedCrossRefGoogle Scholar
  20. 20.
    Janus A, Robak T, Smolewski P (2005) The mammalian target of rapamycin (mTOR) kinase pathway. It’s role in tumourgenesis and targeted antitumour therapy. Cell Mol Biol Lett 10:479–498PubMedGoogle Scholar
  21. 21.
    Wiederrecht GJ, Sabers CJ, Brunn GJ et al (1995) Mechanism of action of rapamycin: new insights into the regulation of G1-phase progression in eukaryotic cells. Prog Cell Cycle Res 1:53–71PubMedGoogle Scholar
  22. 22.
    Neshat MS, Mellinghoff IK, Tran C et al (2001) Enhanced sensitivity of PTEN-deficient tumors to inhibition of FRAP/mTOR. Proc Natl Acad Sci USA 98:10314–10319PubMedCrossRefGoogle Scholar
  23. 23.
    Mondesire W, Jian W, Zhang H et al (2004) Targeting mammalian target of rapamycin synergistically enhances chemotherapy-induced cytotoxicity in breast cancer cells. Clin Cancer Res 10:7031–7042PubMedCrossRefGoogle Scholar
  24. 24.
    Harada H, Andersen J, Mann M et al (2001) p70S6 kinase signals cell survival as well as growth, inactivating the pro-apoptotic molecule BAD. Porc Natl Acad Sci USA 98:9666–9670CrossRefGoogle Scholar
  25. 25.
    Le X, Hittelman W, Liu J et al (2003) Paclitaxel induces inactivation of p70S6 kinase and phosphorylation of The421 and Ser424 via multiple signaling pathways in mitosis. Oncogene 22:484–497PubMedCrossRefGoogle Scholar
  26. 26.
    Wan X, Helman L (2002) Effect of insulin-like growth factor II on protecting myoblast cells against cisplatin-induced apoptosis through p70S6 kinase pathway. Neoplasia 4:400–408PubMedCrossRefGoogle Scholar
  27. 27.
    Yamamoto K, Ichijo H, Korsmeyer S (1999) BCL-2 is phosphorylated and inactivated by an ASK1/Jun N-Terminal protein kinase pathway normally activated at G2/M. Mol Cell Biol 19:8469–8478PubMedGoogle Scholar
  28. 28.
    Floros K, Thomadaki H, Katsaros N et al (2004) mRNA expression analysis of a variety of apoptosis-related genes, including the novel gene of the BCL2-family, BCL2L12, in HL-60 leukemia cells after treatment with carboplatin and doxorubicin. J Biol Chem 385:1099–1103CrossRefGoogle Scholar
  29. 29.
    Raymond E, Alexandre J, Faivre S et al (2002) Dosage adjustment and pharmacokinetic profile of irinotecan in cancer patients with hepatic dysfunction. J Clin Oncol 20:4303–4012PubMedCrossRefGoogle Scholar
  30. 30.
    Ratain MJ, Napoli KL, Knightley Moshier K et al (2007) A phase 1b study of oral rapamycin (sirolimus) in patients with advanced malignancies. J Clin Oncol 25:3510Google Scholar
  31. 31.
    Aguirre D, Boya P, Bellet D et al (2004) Bcl-2 and CCND1/CDK4 expression levels predict the cellular effects of mTOR inhibitors in human ovarian carcinoma. Apoptosis 6:797–805CrossRefGoogle Scholar
  32. 32.
    Noh W, Mondesire W, Peng J et al (2004) Determinants of Rapamycin sensitivity in breast cancer cells. Clin Cancer Res 10:1013–1023PubMedCrossRefGoogle Scholar
  33. 33.
    Wu C, Wangpaichitr M, Feun L et al (2005) Overcoming cispaltin resistance by mTOR inhibitor in lung cancer. Molecular Cancer 4:1–10CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Nasredine Aissat
    • 1
  • Christophe Le Tourneau
    • 1
  • Aïda Ghoul
    • 1
  • Maria Serova
    • 1
  • Ivan Bieche
    • 2
  • François Lokiec
    • 3
  • Eric Raymond
    • 1
  • Sandrine Faivre
    • 1
    • 4
  1. 1.Department of Experimental Pharmacology (RayLab) and Medical OncologyBeaujon University HospitalClichyFrance
  2. 2.Laboratory of Molecular GeneticsBeaujon University HospitalClichyFrance
  3. 3.Department of Clinical PharmacologyCentre René HugueninSaint-CloudFrance
  4. 4.Service Inter-Hospitalier de Cancérologie Bichat-BeaujonClichyFrance

Personalised recommendations