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Combined antitumor effect of γ-secretase inhibitor and ABT-737 in Notch-expressing non-small cell lung cancer

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Abstract

Background

Inhibition of Notch by γ-secretase inhibitor (GSI) has been shown to have an antitumor effect in Notch-expressing non-small cell lung cancer (NSCLC) and to induce apoptosis through modulation of Bcl-2 family proteins. In particular, Bim, a BH3-only member of the Bcl-2 family of proteins, has an important role in the induction of apoptosis in NSCLC when cells are treated with GSI. ABT-737, a BH3-only mimetic, targets the pro-survival Bcl-2 family and also induces apoptosis.

Methods

The Notch-expressing NSCLC cell lines H460, A549, H1793, and HCC2429 were used. The combined antitumor effect of GSI and ABT-737 was evaluated using the MTT proliferation assay in vitro and in xenograft mouse models. The expression of the Notch pathway and Bcl-2 family was analyzed using Western blotting analysis when cells were treated with a single drug treatment or a combination treatment.

Results

GSI XX or ABT-737 alone inhibited cell proliferation in a dose-dependent manner, and combination drug treatment showed a synergistic antitumor effect in vitro. In vivo, this drug combination significantly suppressed tumor proliferation compared to the single drug treatment. Phospho-Bcl-2 was downregulated and Bax was upregulated by both the single and combination drug treatments. Bim was induced by a single drug treatment and was enhanced by combination treatment. Combination treatment-induced apoptosis was decreased by Bim inhibition, suggesting that the antitumor effect of the drug combination was dependent on Bim.

Conclusion

Based on our data, we propose that the combination treatment is a promising strategy for NSCLC therapy.

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References

  1. Maemondo M, Inoue A, Kobayashi K et al (2010) Gefitinib or chemotherapy for non-small-cell lung cancer with mutated EGFR. N Engl J Med 362:2380–2388

    Article  CAS  PubMed  Google Scholar 

  2. Sandler A, Gray R, Perry MC et al (2006) Paclitaxel-carboplatin alone or with bevacizumab for non-small-cell lung cancer. N Engl J Med 355:2542–2550

    Article  CAS  PubMed  Google Scholar 

  3. Allenspach EJ, Maillard I, Aster JC et al (2002) Notch signaling in cancer. Cancer Biol Ther 1:466–476

    Article  PubMed  Google Scholar 

  4. Iso T, Kedes L, Hamamori Y (2003) HES and HERP families: multiple effectors of the Notch signaling pathway. J Cell Physiol 194:237–255

    Article  CAS  PubMed  Google Scholar 

  5. Das I, Craig C, Funahashi Y et al (2004) Notch oncoproteins depend on gamma-secretase/presenilin activity for processing and function. J Biol Chem 279:30771–33080

    Article  CAS  PubMed  Google Scholar 

  6. Curry CL, Reed LL, Broude E et al (2007) Notch inhibition in Kaposi’s sarcoma tumor cells leads to mitotic catastrophe through nuclear factor-kappaB signaling. Mol Cancer Ther 6:1983–1992

    Article  CAS  PubMed  Google Scholar 

  7. Duechler M, Shehata M, Schwarzmeier JD et al (2005) Induction of apoptosis by proteasome inhibitors in B-CLL cells is associated with downregulation of CD23 and inactivation of Notch2. Leukemia 19:260–267

    Article  CAS  PubMed  Google Scholar 

  8. Reedijk M, Odorcic S, Chang L et al (2005) High-level coexpression of JAG1 and NOTCH1 is observed in human breast cancer and is associated with poor overall survival. Cancer Res 65:8530–8537

    Article  CAS  PubMed  Google Scholar 

  9. Konishi J, Kawaguchi KS, Vo H et al (2007) Gamma-secretase inhibitor prevents Notch3 activation and reduces proliferation in human lung cancers. Cancer Res 67:8051–8057

    Article  CAS  PubMed  Google Scholar 

  10. Hengatner MO (2000) The biochemistry of apoptosis. Nature (Lond) 407:770–776

    Article  Google Scholar 

  11. Cory S, Huang DC, Adamas JM (2003) The Bcl-2 family: roles in cell survival and oncogenesis. Oncogene 22:8590–8607

    Article  CAS  PubMed  Google Scholar 

  12. Adams JM, Cory S (2007) The Bcl-2 apoptotic switch in cancer development and therapy. Oncogene 26:1324–1337

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Petros AM, Olejniczak ET, Fesik SW (2004) Structural biology of the Bcl-2 family of proteins. Biochem Biophys Acta 1644:83–94

    Article  CAS  PubMed  Google Scholar 

  14. Willis SN, Fletcher JI, Kaufmann T et al (2007) Apoptosis initiated when BH3 ligands engage multiple Bcl-2 homologs, not Bax or Bak. Science 315:856–859

    Article  CAS  PubMed  Google Scholar 

  15. Konishi J, Yi F, Chen X et al (2010) Notch3 cooperates with the EGFR pathway to modulate apoptosis through the induction of Bim. Oncogene 29:589–596

    Article  CAS  PubMed  Google Scholar 

  16. Oltersdorf T, Elmore SW, Shoemaker AR et al (2005) An inhibitor of Bcl-2 family proteins induces regression of solid tumours. Nature (Lond) 435:677–681

    Article  CAS  Google Scholar 

  17. Chauhan D, Velankar M, Brahmandam M et al (2007) A novel Bcl-2/Bcl-X(L)/Bcl-w inhibitor ABT-737 as therapy in multiple myeloma. Oncogene 26:2374–2380

    Article  CAS  PubMed  Google Scholar 

  18. Tahir SK, Yang X, Anderson MG et al (2007) Influence of Bcl-2 family members on the cellular response of small-cell lung cancer cell lines to ABT-737. Cancer Res 67:1176–1183

    Article  CAS  PubMed  Google Scholar 

  19. Tagscherer KE, Fassl A, Campos B et al (2008) Apoptosis-based treatment of glioblastomas with ABT-737, a novel small molecule inhibitor of Bcl-2 family proteins. Oncogene 27:6646–6656

    Article  CAS  PubMed  Google Scholar 

  20. Li M, Chen F, Clifton N et al (2010) Combined inhibition of Notch signaling and Bcl-2/Bcl-xL results in synergistic antimyeloma effect. Mol Cancer Ther 9:3200–3209

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Séveno C, Loussouarn D, Bréchet S et al (2012) γ-Secretase inhibition promotes cell death, Noxa upregulation, and sensitization to BH3 mimetic ABT-737 in human breast cancer cells. Breast Cancer Res 14:R96

    Article  PubMed  PubMed Central  Google Scholar 

  22. Dang TP, Gazdar AF, Virmani AK et al (2000) Chromosome 19 translocation, overexpression of Notch3, and human lung cancer. J Natl Cancer Inst 92:1355–1357

    Article  CAS  PubMed  Google Scholar 

  23. Chou TC (2010) Drug combination studies and their synergy quantification using the Chou–Talalay method. Cancer Res 70:440–446

    Article  CAS  PubMed  Google Scholar 

  24. Dalrymple S, Antony L, Xu Y et al (2005) Role of Notch-1 and E-cadherin in the differential response to calcium in culturing normal versus malignant prostate cells. Cancer Res 65:9269–9279

    Article  CAS  PubMed  Google Scholar 

  25. Costa DB, Halmos B, Kumar A et al (2007) BIM mediates EGFR tyrosine kinase inhibitor-induced apoptosis in lung cancers with oncogenic EGFR mutations. PloS Med 10:1669–1679

    Google Scholar 

  26. Yoo AS, Bais C, Greenwald I (2004) Crosstalk between the EGFR and LIN-12/Notch pathways in C. elegans vulval development. Science 303:663–666

    Article  CAS  PubMed  Google Scholar 

  27. Lammich S, Okochi M, Takeda M et al (2002) Presenilin-dependent intramembrane proteolysis of CD44 leads to the liberation of its intracellular domain and the secretion of an Abeta-like peptide. J Biol Chem 277:44754–44759

    Article  CAS  PubMed  Google Scholar 

  28. Ni CY, Murphy MP, Golde TE et al (2001) Gamma-secretase cleavage and nuclear localization of ErbB-4 receptor tyrosine kinase. Science 294:2179–2181

    Article  CAS  PubMed  Google Scholar 

  29. Mizugaki H, Sakakibara-Konishi J, Ikezawa Y et al (2012) γ-Secretase inhibitor enhances antitumour effect of radiation in Notch-expressing lung cancer. Br J Cancer 106:1953–1959

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Will B, Siddiqi T, Jordà MA et al (2010) Apoptosis induced by JAK2 inhibition is mediated by Bim and enhanced by the BH3 mimetic ABT-737 in JAK2 mutant human erythroid cells. Blood 115:2901–2919

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Wang H, Yang YB, Shen HM et al (2012) ABT-737 induces Bim expression via JNK signaling pathway and its effect on the radiation sensitivity of HeLa cells. PLoS One 7:e52483

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Goldsmith KC, Gross M, Peirce S et al (2012) Mitochondrial Bcl-2 family dynamics define therapy response and resistance in neuroblastoma. Cancer Res 72:2565–2577

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Lucas KM, Mohana-Kumaran N, Lau D et al (2012) Modulation of NOXA and MCL-1 as a strategy for sensitizing melanoma cells to the BH3-mimetic ABT-737. Clin Cancer Res 18:783–795

    Article  CAS  PubMed  Google Scholar 

  34. Sriuranpong V, Borges MW, Ravi RK et al (2001) Notch signaling induces cell cycle arrest in small cell lung cancer cells. Cancer Res 61:3200–3205

    CAS  PubMed  Google Scholar 

  35. Hassan WA, Yoshida R, Kudoh S et al (2014) Notch1 controls cell invasion and metastasis in small cell lung carcinoma cell lines. Lung Cancer 86:304–310

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

We thank M. Maeda (First Department of Medicine, Hokkaido University School of Medicine) for providing special support and experimental assistance during this study.

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Authors and Affiliations

  1. First Department of Medicine, Hokkaido University School of Medicine, North 15, West 7, Kita-ku, Sapporo, 060-8638, Japan

    Jun Sakakibara-Konishi, Yasuyuki Ikezawa, Satoshi Oizumi, Junko Kikuchi, Eiki Kikuchi, Hidenori Mizugaki & Masaharu Nishimura

  2. Department of Medical Oncology, Hokkaido University Graduate School of Medicine, Sapporo, Japan

    Ichiro Kinoshita & Hirotoshi Dosaka-Akita

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  1. Jun Sakakibara-Konishi
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  2. Yasuyuki Ikezawa
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Correspondence to Jun Sakakibara-Konishi.

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10147_2016_1060_MOESM1_ESM.pptx

Supplementary Fig. S1: The expression of BH3-only proteins in control or drug-treated NSCLC cell lines was evaluated by Western blotting. No significant change in expression was observed between control and treated cells. The expression of Bim and Noxa is presented in Fig. 3C. (PPTX 266 kb)

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Sakakibara-Konishi, J., Ikezawa, Y., Oizumi, S. et al. Combined antitumor effect of γ-secretase inhibitor and ABT-737 in Notch-expressing non-small cell lung cancer. Int J Clin Oncol 22, 257–268 (2017). https://doi.org/10.1007/s10147-016-1060-3

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  • DOI: https://doi.org/10.1007/s10147-016-1060-3

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