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Vol. 85, No. 1, 2007
Issue release date: April 2007

The Novel mTOR Inhibitor RAD001 (Everolimus) Induces Antiproliferative Effects in Human Pancreatic Neuroendocrine Tumor Cells

Zitzmann K. · De Toni E.N. · Brand S. · Göke B. · Meinecke J. · Spöttl G. · Meyer H.H.D. · Auernhammer C.J.
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Background/Aim: Tumors exhibiting constitutively activated PI(3)K/Akt/mTOR signaling are hypersensitive to mTOR inhibitors such as RAD001 (everolimus) which is presently being investigated in clinical phase II trials in various tumor entities, including neuroendocrine tumors (NETs). However, no preclinical data about the effects of RAD001 on NET cells have been published. In this study, we aimed to evaluate the effects of RAD001 on BON cells, a human pancreatic NET cell line that exhibits constitutively activated PI(3)K/Akt/mTOR signaling. Methods: BON cells were treated with different concentrations of RAD001 to analyze its effect on cell growth using proliferation assays. Apoptosis was examined by Western blot analysis of caspase-3/PARP cleavage and by FACS analysis of DNA fragmentation. Results: RAD001 potently inhibited BON cell growth in a dose-dependent manner which was dependent on the serum concentration in the medium. RAD001-induced growth inhibition involved G0/G1-phase arrest as well as induction of apoptosis. Conclusion: In summary, our data demonstrate antiproliferative and apoptotic effects of RAD001 in NET cells in vitro supporting its clinical use in current phase II trials in NET patients.

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  1. Shaw RJ, Cantley LC: Ras, PI(3)K and mTOR signalling controls tumour cell growth. Nature 2006;441:424–430.
  2. Wullschleger S, Loewith R, Hall MN: TOR signaling in growth and metabolism. Cell 2006;124:471–484.
  3. O’Reilly KE, Rojo F, She QB, Solit D, Mills GB, Smith D, Lane H, Hofmann F, Hicklin DJ, Ludwig DL, Baselga J, Rosen N: mTOR inhibition induces upstream receptor tyrosine kinase signaling and activates Akt. Cancer Res 2006;66:1500–1508.
  4. Neshat MS, Mellinghoff IK, Tran C, Stiles B, Thomas G, Petersen R, Frost P, Gibbons JJ, Wu H, Sawyers CL: Enhanced sensitivity of PTEN-deficient tumors to inhibition of FRAP/mTOR. Proc Natl Acad Sci USA 2001;98:10314–10319.
  5. Wendel HG, Malina A, Zhao Z, Zender L, Kogan SC, Cordon-Cardo C, Pelletier J, Lowe SW: Determinants of sensitivity and resistance to rapamycin-chemotherapy drug combinations in vivo. Cancer Res 2006;66:7639–7646.
  6. Faivre S, Kroemer G, Raymond E: Current development of mTOR inhibitors as anticancer agents. Nat Rev Drug Discov 2006;5:671–688.
  7. Beuvink I, Boulay A, Fumagalli S, Zilbermann F, Ruetz S, O’Reilly T, Natt F, Hall J, Lane HA, Thomas G: The mTOR inhibitor RAD001 sensitizes tumor cells to DNA-damaged induced apoptosis through inhibition of p21 translation. Cell 2005;120:747–759.
  8. Boulay A, Rudloff J, Ye J, Zumstein-Mecker S, O’Reilly T, Evans DB, Chen S, Lane HA: Dual inhibition of mTOR and estrogen receptor signaling in vitro induces cell death in models of breast cancer. Clin Cancer Res 2005;11:5319–5328.
  9. Boulay A, Zumstein-Mecker S, Stephan C, Beuvink I, Zilbermann F, Haller R, Tobler S, Heusser C, O’Reilly T, Stolz B, Marti A, Thomas G, Lane HA: Antitumor efficacy of intermittent treatment schedules with the rapamycin derivative RAD001 correlates with prolonged inactivation of ribosomal protein S6 kinase 1 in peripheral blood mononuclear cells, Cancer Res 2004;64:252–261.
  10. Wanner K, Hipp S, Oelsner M, Ringshausen I, Bogner C, Peschel C, Decker T: Mammalian target of rapamycin inhibition induces cell cycle arrest in diffuse large B cell lymphoma (DLBCL) cells and sensitises DLBCL cells to rituximab. Br J Haematol 2006;134:475–484.
  11. Albert JM, Kim KW, Cao C, Lu B: Targeting the Akt/mammalian target of rapamycin pathway for radiosensitization of breast cancer. Mol Cancer Ther 2006;5:1183–1189.
  12. Cao C, Subhawong T, Albert Jm, Kim KW, Geng L, Sekhar KR, Gi YJ, Lu B: Inhibition of mammalian target of rapamycin or apoptotic pathway induces autophagy and radiosensitizes PTEN null prostate cancer cells. Cancer Res 2006;66:10040–10047.
  13. Maggard MA, O’Connell JB, Ko CY: Updated population-based review of carcinoid tumors. Ann Surg 2004;240:117–122.
  14. Modlin IM, Lye KD, Kidd M: A 5-decade analysis of 13,715 carcinoid tumors. Cancer 2003;97:934–959.
  15. Kaltsas GA, Besser GM, Grossman AB: The diagnosis and medical management of advanced neuroendocrine tumors. Endocr Rev 2004;25:458–511.
  16. Mignon M: Natural history of neuroendocrine enteropancreatic tumors. Digestion 2000;62(suppl 1):51–58.
  17. Arnold R, Rinke A, Schmidt C, Hofbauer L: Endocrine tumours of the gastrointestinal tract: Chemotherapy Best Pract Res Clin Gastroenterol 2005;19:649–656.
  18. Oberg K: Management of neuroendocrine tumours. Ann Oncol 2004;15(suppl 4):iv293–iv298.

    External Resources

  19. Ramage JK, Davies AH, Ardill J, Bax N, Caplin M, Grossman A, Hawkins R, McNicol AM, Reed N, Sutton R, Thakker R, Aylwin S, Breen D, Britton K, Buchanan K, Corrie P, Gillams A, Lewington V, McCance D, Meeran K, Watkinson A: Guidelines for the management of gastroenteropancreatic neuroendocrine (including carcinoid) tumours. Gut 2005;54(suppl 4):iv1–iv16.
  20. Shah T, Hochhauser D, Frow R, Quaglia A, Dhillon AP, Caplin ME: Epidermal growth factor receptor expression and activation in neuroendocrine tumours. J Neuroendocrinol 2006;18:355–360.
  21. Wang L, Ignat A, Axiotis CA: Differential expression of the PTEN tumor suppressor protein in fetal and adult neuroendocrine tissues and tumors: progressive loss of PTEN expression in poorly differentiated neuroendocrine neoplasms. Appl Immunohistochem Mol Morphol 2002;10:139–146.
  22. Jeng YJ, Townsend CM Jr, Nagasawa S, Chuo S, Kern K, Yanaihara N, Ferrar RS, Hill FL, Thompson JC, Greeley GH Jr: Regulation of pancreastatin release from a human pancreatic carcinoid cell line in vitro. Endocrinology 1991;128:220–225.
  23. Evers BM, Ishizuka J, Townsend CM Jr, Thompson JC: The human carcinoid cell line, BON. A model system for the study of carcinoid tumors. Ann NY Acad Sci 1994;733:393–406.
  24. von Wichert G, Jehle PM, Hoeflich A, Koschnick S, Dralle H, Wolf E, Wiedenmann B, Boehm BO, Adler G, Seufferlein T: Insulin-like growth factor-I is an autocrine regulator of chromogranin A secretion and growth in human neuroendocrine tumor cells. Cancer Res 2000;60:4573–4581.
  25. Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C: A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 1991;139:271–279.
  26. Vlotides G, Sorensen AS, Kopp F, Zitzmann K, Cengic N, Brand S, Zachoval R, Auernhammer CJ: SOCS-1 and SOCS-3 inhibit IFN-alpha-induced expression of the antiviral proteins 2,5-OAS and MxA. Biochem Biophys Res Commun 2004;320:1007–1014.
  27. Auernhammer CJ, Dorn F, Vlotides G, Hengge S, Kopp FB, Spoettl G, Cengic N, Engelhardt D, Weber MM: The oncostatin M receptor/gp130 ligand murine oncostatin M induces apoptosis in adrenocortical Y-1 tumor cells. J Endocrinol 2004;180:479–486.
  28. Sun SY, Rosenberg LM, Wang X, Zhou Z, Yue P, Fu H, Khuri FR: Activation of Akt and eIF4E survival pathways by rapamycin-mediated mammalian target of rapamycin inhibition. Cancer Res 2005;65:7052–7058.
  29. Luo J, Manning BD, Cantley LC: Targeting the PI3K-Akt pathway in human cancer: rationale and promise. Cancer Cell 2003;4:257–262.
  30. Sawyers CL: Will kinase inhibitors have a dark side? N Engl J Med 2006;355:313–315.
  31. Kirchner GI, Meier-Wiedenbach I, Manns MP: Clinical pharmacokinetics of everolimus. Clin Pharmacokinet 2004;43:83–95.
  32. Vignot S, Faivre S, Aguirre D, Raymond E: mTOR-targeted therapy of cancer with rapamycin derivatives. Ann Oncol 2005;16:525–537.
  33. Yee KW, Zeng Z, Konopleva M, Verstovsek S, Ravandi F, Ferrajoli A, Thomas D, Wierda W, Apostolidou E, Albitar M, O’Brien S, Andreeff M, Giles FJ: Phase I/II study of the mammalian target of rapamycin inhibitor everolimus (RAD001) in patients with relapsed or refractory hematologic malignancies. Clin Cancer Res 2006;12:5165–5173.
  34. Grewe M, Gansauge F, Schmid RM, Adler G, Seufferlein T: Regulation of cell growth and cyclin D1 expression by the constitutively active FRAP-p70s6K pathway in human pancreatic cancer cells. Cancer Res 1999;59:3581–3587.
  35. Hashemolhosseini S, Nagamine Y, Morley SJ, Desrivieres S, Mercep L, Ferrari S: Rapamycin inhibition of the G1 to S transition is mediated by effects on cyclin D1 mRNA and protein stability. J Biol Chem 1998;273:14424–14429.
  36. Kawamata S, Sakaida H, Hori T, Maeda M, Uchiyama T: The upregulation of p27Kip1 by rapamycin results in G1 arrest in exponentially growing T-cell lines. Blood 1998;91:561–569.
  37. Hosoi H, Dilling MB, Shikata T, Liu LN, Shu L, Ashmun RA, Germain GS, Abraham RT, Houghton PJ: Rapamycin causes poorly reversible inhibition of mTOR and induces p53-independent apoptosis in human rhabdomyosarcoma cells. Cancer Res 1999;59:886–894.
  38. Yao JC, Phan AT, Chang DZ, Jacobs C, Mares JE, Rashid A, Meric-Bernstam F: Phase II study of RAD001 (everolimus) and depot octreotide (Sandostatin LAR) in patients with advanced low grade neuroendocrine carcinoma (LGNET). ASCO Annual Meeting Proceedings Part I. J Clin Oncol 2006;24:4042.
  39. Bousquet C, Guillermet-Guibert J, Saint-Laurent N, Archer-Lahlou E, Lopez F, Fanjul M, Ferrand A, Fourmy D, Pichereaux C, Monsarrat B, Pradayrol L, Esteve JP, Susini C: Direct binding of p85 to sst2 somatostatin receptor reveals a novel mechanism for inhibiting PI3K pathway. EMBO J 2006;25:3943–3954.

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