Medical Oncology

, Volume 27, Issue 3, pp 1017–1022 | Cite as

Inhibition of Notch3 enhances sensitivity to gemcitabine in pancreatic cancer through an inactivation of PI3K/Akt-dependent pathway

  • Jun Yao
  • Cuijuan Qian
Original Paper


Notch3 is one of the four Notch receptors identified in mammal, but its role in human pancreatic cancer remains poorly characterized. In this study, we sought to determine the effect of suppressing Notch3 expression on the chemosensitivity to gemcitabine in human pancreatic cancer cell lines BxPC-3 and PANC-1. RNA interference was used to suppress Notch3 expression. Gemcitabine-induced cytotoxicity was determined by MTT. Cell apoptosis was measured by flow cytometry. Caspase 3 activity was assayed using a Caspase Fluorescent Assay Kit. The effect of Notch3-specific siRNA on PI3K/Akt activity was also quantified. Notch3-specific siRNA suppressed Notch3 expression, and furthermore increased gemcitabine-induced, caspase-mediated apoptosis. The suppression of Notch3 expression decreased the average IC50 in BxPC-3 and PANC-1 cells treated with gemcitabine. PI3K/Akt activity was decreased by the suppression of Notch3 expression. Taken together, these data demonstrated that Notch3 is a potential therapeutic target for pancreatic cancer, and PI3K/Akt is a key signaling component by which activation of the Notch3 signal transduction pathway protects pancreatic cancer cells from chemotherapy-induced cell death.


Pancreatic cancer Notch3 Caspase PI3K/Akt Gemcitabine 


  1. 1.
    Fryer RA, Galustian C, Dalgelish AG. Recent advances and developments in treatment strategies against pancreatic cancer. Curr Clin Pharmacol. 2009;4(2):102–12.CrossRefPubMedGoogle Scholar
  2. 2.
    Jonckheere N, et al. Tumour growth and resistance to gemcitabine of pancreatic cancer cells are decreased by AP-2alpha overexpression. Br J Cancer. 2009;101(4):637–44.CrossRefPubMedGoogle Scholar
  3. 3.
    Nickoloff BJ, Osborne BA, Miele L. Notch signaling as a therapeutic target in cancer: a new approach to the development of cell fate modifying agents. Oncogene. 2003;22(42):6598–608.CrossRefPubMedGoogle Scholar
  4. 4.
    Miyamoto A, Lau R, Hein PW, Shipley JM, Weinmaster G. Microfibrillar proteins MAGP-1 and MAGP-2 induce Notch1 extracellular domain dissociation and receptor activation. J Biol Chem. 2006;281(15):10089–97.CrossRefPubMedGoogle Scholar
  5. 5.
    Radtke F, Wilson A, Mancini SJ, MacDonald HR. Notch regulation of lymphocyte development and function. Nat Immunol. 2004;5(3):247–53.CrossRefPubMedGoogle Scholar
  6. 6.
    Maillard I, Adler SH, Pear WS. Notch and the immune system. 1. Immunity. 2003;19(6):781–91.CrossRefPubMedGoogle Scholar
  7. 7.
    Maillard I, Fang T, Pear WS. Regulation of lymphoid development, differentiation, and function by the Notch pathway. Annu Rev Immunol. 2005;23:945–74.CrossRefPubMedGoogle Scholar
  8. 8.
    Real PJ, Ferrando AA. NOTCH inhibition and glucocorticoid therapy in T-cell acute lymphoblastic leukemia. Leukemia. 2009;23(8):1374–7.CrossRefPubMedGoogle Scholar
  9. 9.
    Roy M, Pear WS, Aster JC. The multifaceted role of Notch in cancer. Curr Opin Genet Dev. 2007;17(1):52–9.CrossRefPubMedGoogle Scholar
  10. 10.
    Dotto GP. Notch tumor suppressor function. Oncogene. 2008;27(38):5115–23.CrossRefPubMedGoogle Scholar
  11. 11.
    Baek SH, et al. Zinc-induced downregulation of Notch signaling is associated with cytoplasmic retention of Notch1-IC and RBP-Jk via PI3k-Akt signaling pathway. Cancer Lett. 2007;255(1):117–26.CrossRefPubMedGoogle Scholar
  12. 12.
    Meurette O, et al. Notch activation induces Akt signaling via an autocrine loop to prevent apoptosis in breast epithelial cells. Cancer Res. 2009;69(12):5015–22.CrossRefPubMedGoogle Scholar
  13. 13.
    Kurreck J. RNA interference: from basic research to therapeutic applications. Angew Chem Int Ed Engl. 2009;48(8):1378–98.CrossRefPubMedGoogle Scholar
  14. 14.
    Hu Y, et al. Inhibition of hypoxia-inducible factor-1 function enhances the sensitivity of multiple myeloma cells to melphalan. Mol Cancer Ther. 2009;8(8):2329–38.CrossRefPubMedGoogle Scholar
  15. 15.
    Pauwels B, et al. The role of apoptotic cell death in the radiosensitising effect of gemcitabine. Br J Cancer. 2009;101(4):628–36.CrossRefPubMedGoogle Scholar
  16. 16.
    Zhou H, Li XM, Meinkoth J, Pittman RN. Akt regulates cell survival and apoptosis at a postmitochondrial level. J Cell Biol. 2000;151(3):483–94.CrossRefPubMedGoogle Scholar
  17. 17.
    Simon PO Jr, et al. Targeting AKT with the proapoptotic peptide, TAT-CTMP: a novel strategy for the treatment of human pancreatic adenocarcinoma. Int J Cancer. 2009;125(4):942–51.CrossRefPubMedGoogle Scholar
  18. 18.
    Gramantieri L, et al. Aberrant Notch3 and Notch4 expression in human hepatocellular carcinoma. Liver Int. 2007;27(7):997–1007.CrossRefPubMedGoogle Scholar
  19. 19.
    Doucas H, et al. Expression of nuclear Notch3 in pancreatic adenocarcinomas is associated with adverse clinical features, and correlates with the expression of STAT3 and phosphorylated Akt. J Surg Oncol. 2008;97(1):63–8.CrossRefPubMedGoogle Scholar
  20. 20.
    Dang L, et al. Notch3 signaling initiates choroid plexus tumor formation. Oncogene. 2006;25(3):487–91.PubMedGoogle Scholar
  21. 21.
    Akiyoshi T, et al. Gamma-secretase inhibitors enhance taxane-induced mitotic arrest and apoptosis in colon cancer cells. Gastroenterology. 2008;134(1):131–44.CrossRefPubMedGoogle Scholar
  22. 22.
    Gazzaniga P, et al. Gemcitabine-induced apoptosis in 5637 cell line: an in vitro model for high-risk superficial bladder cancer. Anticancer Drugs. 2007;18(2):179–85.CrossRefPubMedGoogle Scholar
  23. 23.
    Nefedova Y, Sullivan DM, Bolick SC, Dalton WS, Gabrilovich DI. Inhibition of Notch signaling induces apoptosis of myeloma cells and enhances sensitivity to chemotherapy. Blood. 2008;111(4):2220–9.CrossRefPubMedGoogle Scholar
  24. 24.
    Tassone P, et al. Zoledronic acid induces antiproliferative and apoptotic effects in human pancreatic cancer cells in vitro. Br J Cancer. 2003;88(12):1971–8.CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2009

Authors and Affiliations

  1. 1.School of MedicineTaizhou UniversityTaizhouChina
  2. 2.Department of GastroenterologyTaizhou Municipal HospitalTaizhouChina

Personalised recommendations