Skip to main content

γ-Secretase inhibitor, DAPT inhibits self-renewal and stemness maintenance of ovarian cancer stem-like cells in vitro

Chinese Journal of Cancer Research volume 23, Article number: 140 (2011) Cite this article

Abstract

Objective

The Notch signaling pathway plays an important role in the stem cell signaling network and contributes to tumorigenesis. However, the functions of Notch signaling in ovarian cancer stem cells (OCSCs) are not well understood. We aimed to investigate the effects of Notch blockade on self-renewal and stemness maintenance of OCSCs.

Methods

Ovarian cancer stem-like cells were enriched from ovarian cancer cell lines in serum-free medium. A γ-secretase inhibitor, (DAPT), was used to block Notch signaling. MTT assays were performed to assess self-renewal and proliferation inhibition, flow cytometry was performed to analyze cell surface marker and immunofluorescence, Western Blot and Real-time RT-PCR assays were performed to detect Oct4 and Sox2 protein and mRNA expression of the Ovarian cancer stem-like cells treated with DAPT.

Results

Notch blockade markedly inhibits self-renewal and proliferation of ovarian cancer stem-like cells, significantly downregulates the expression of OCSCs-specific surface markers, and reduces protein and mRNA expression of Oct4 and Sox2 in OCSC-like cells.

Conclusion

Our results suggest that Notch signaling is not only critical for the self-renewal and proliferation of OCSCs, but also for the stemness maintenance of OCSCs. The γ-secretase inhibitor is a promising treatment targeting OCSCs.

This is a preview of subscription content, access via your institution.

References

  1. Pecorelli S, Favalli G, Zigliani L, et al. Cancer in women. Int J Gynaecol Obstet 2003; 82: 369–379.

    PubMed  Article  CAS  Google Scholar 

  2. Dick JE. Stem cell concepts renew cancer research. Blood 2008; 112: 4793–4807.

    PubMed  Article  CAS  Google Scholar 

  3. Clarke MF, Dick JE, Dirks PB, et al. Cancer Stem Cells—Perspectives on Current Status and Future Directions: AACR Workshop on Cancer Stem Cells. Cancer Res 2006; 66: 9339–9344.

    PubMed  Article  CAS  Google Scholar 

  4. Zhang S, Balch C, Chan MW, et al: Identification and Characterization of Ovarian Cancer-Initiating Cells from Primary Human Tumors. Cancer Res 2008; 68: 4311–4320.

    PubMed  Article  CAS  Google Scholar 

  5. Ailles LE, Weissman IL. Cancer stem cells in solid tumors. Curr Opin Biotechnol 2007; 18: 460–466.

    PubMed  Article  CAS  Google Scholar 

  6. Bapat SA, Mali AM, Koppikar CB, et al. Stem and progenitor-like cells contribute to the aggressive behavior of human epithelial ovarian cancer. Cancer Res 2005; 65: 3025–3029.

    PubMed  CAS  Google Scholar 

  7. Burleson KM, Casey RC, Skubitz KM, et al. Ovarian carcinoma ascites spheroids adhere to extracellular matrix components and mesothelial cell monolayers. Gynecol Oncol 2004; 93: 170–181.

    PubMed  Article  CAS  Google Scholar 

  8. Curley MD, Therrien VA, Cummings CL, et al. CD133 expression defines a tumor initiating cell population in primary human ovarian cancer. Stem Cells 2009; 27: 2875–2883.

    PubMed  CAS  Google Scholar 

  9. Wani AA, Sharma N, Shouche YS, et al. Nuclear-mitochondrial genomic profiling reveals a pattern of evolution in epithelial ovarian tumor stem cells. Oncogene 2006; 25: 6336–6344.

    PubMed  Article  CAS  Google Scholar 

  10. Szotek PP, Pieretti-Vanmarcke R, Masiakos PT, et al. Ovarian cancer side population defines cells with stem cell-like characteristics and Mullerian Inhibiting Substance responsiveness. Proc Natl Acad Sci USA 2006; 103: 11154–11159.

    PubMed  Article  CAS  Google Scholar 

  11. Mimeault M, Hauke R, Mehta PP, et al. Recent advances in cancer stem/progenitor celll research: therapeutic implications for overcoming resistance to the most aggressive cancers. J Cell Mol Med 2007; 11: 981–1011.

    PubMed  Article  CAS  Google Scholar 

  12. Leong KG, Karsan A. Recent insight into the role of Notch signaling in tumorigenesis. Blood 2006; 107: 2223–2233.

    PubMed  Article  CAS  Google Scholar 

  13. AI-Hajj M, Clarke MF. Self-renewal and solid tumor stem cells. Oncogene 2004; 23: 7274–7282.

    Article  Google Scholar 

  14. Artavanis-Tsakonas S, Rand MD, Lake RJ. Notch Signaling: Cell Fate Control and Signal Integration in Development. Science 1999; 284: 770–776.

    PubMed  Article  CAS  Google Scholar 

  15. Androutsellis-Theotokis A, Leker RR, Soldner F, et al. Notch signaling regulates stem cell numbers in vitro and in vivo. Nature 2006; 442: 823–826.

    PubMed  Article  CAS  Google Scholar 

  16. Li JL, Harris AL. Notch signaling from tumor cells: a new mechanism of angiogenesis. Cancer Cell 2005; 8: 1–3.

    PubMed  Article  CAS  Google Scholar 

  17. Hallahan AR, Pritchard JI, Hansen S, et al. The SmoA1 mouse model reveals that notch signaling is critical for the growth and survival of sonic hedgehog-induced medulloblastomas. Cancer Res 2004; 64: 7794–7800.

    PubMed  Article  CAS  Google Scholar 

  18. Sjölund J, Johansson M, Manna S, et al. Suppression of renal cell carcinoma growth by inhibition of Notch signaling in vitro and in vivo. J Clin Invest 2008; 118: 217–228.

    PubMed  Article  Google Scholar 

  19. Kimura K, Satoh K, Kanno A, et al. Activation of Notch signaling in tumorigenesis of experimental pancreatic cancer induced by dimethylbenzanthracene in mice. Cancer Sci 2007; 98: 155–162.

    PubMed  Article  CAS  Google Scholar 

  20. Rappa G, Mercapide J, Anzanello F, et al. Growth of cancer cell lines under stem cell-like conditions has the potential to unveil therapeutic targets. Exp Cell Res 2008; 314: 2110–2122.

    PubMed  Article  CAS  Google Scholar 

  21. Bez A, Corsini E, Curti D, et al. Neurosphere and neurosphere-forming cells: morphological and ultrastructural characterization. Brain Res 2003; 993: 18–29.

    PubMed  Article  CAS  Google Scholar 

  22. Dontu G, Abdallah WM, Foley JM, et al. In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells. Genes Dev 2003; 17: 1253–1270.

    PubMed  Article  CAS  Google Scholar 

  23. Bisson I, Prowse DM. WNT signaling regulates self-renewal and differentiation of prostate cancer cells with stem cell characteristics. Cell Res 2009; 19: 683–697.

    PubMed  Article  CAS  Google Scholar 

  24. Reya T, Morrison SJ, Clarke MF, et al. Stem cells,cancer,and cancer stem cells. Nature 2001; 414: 105–111.

    PubMed  Article  CAS  Google Scholar 

  25. Bray SJ. Notch signaling: a simple pathway becomes complex. Nat Rev Mol Cell Biol 2006; 7: 678–689.

    PubMed  Article  CAS  Google Scholar 

  26. Katoh M, Katoh M. Notch signaling in gastrointestinal tract. Int J Onco 2007; 30: 247–251.

    CAS  Google Scholar 

  27. Shih IeM, Wang TL. Notch Signaling, gamma-Secretase Inhibitors, and Cancer Therapy. Cancer Res 2007; 67: 1879–1882.

    PubMed  Article  CAS  Google Scholar 

  28. Raspollini MR, Amunni G, Villanucci A, et al. c-KIT expression and correlation with chemotherapy resistance in ovarian carcinoma: an immunocytochemical study. Ann Oncol 2004; 15: 594–597.

    PubMed  Article  CAS  Google Scholar 

  29. Saegusa M, Machida D, Hashimura M, et al. CD44 expression in benign, premalignant, and malignant ovarian neoplasms: relation to tumour development and progression. J Pathol 1999; 189: 326–337.

    PubMed  Article  CAS  Google Scholar 

  30. Shmelkov S, Clair RSt, Lyden D, et al. AC133/CD133/Prominin-1. Int J Biochem Cell Biol 2005; 37: 715–719.

    PubMed  Article  CAS  Google Scholar 

  31. Mimeault M, Hauke R, Mehta PP, et al. Recent advances in cancer stem/progenitor cell research: therapeutic implications for overcoming resistance to the most aggressive cancers. J Cell Mol Med 2007; 11: 981–1011.

    PubMed  Article  CAS  Google Scholar 

  32. Baba T, Convery PA, Matsumura N, et al. Epigenetic regulation of CD133 and tumorigenicity of CD133+ ovarian cancer cells. Oncogene 2009; 28: 209–218.

    PubMed  Article  CAS  Google Scholar 

  33. Huangfu D, Osafune K, Maehr R, et al. Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2. Nat Biotechnol 2008; 26: 1269–1275.

    PubMed  Article  CAS  Google Scholar 

  34. Ben-Porath I, Thomson MW, Carey VJ, et al. An embryonic stem cell-like gene expression signature in poorly differentiated aggressive human tumors. Nat Genet 2008; 40: 499–507.

    PubMed  Article  CAS  Google Scholar 

  35. Nicolis SK. Cancer stem cells and “stemness” genes in neuro-oncology. Neurobiol Dis 2007; 25: 217–229.

    PubMed  Article  CAS  Google Scholar 

Download references

Author information

Affiliations

  1. Department of Obstetrics and Gynecology, the First Affiliated Hospital, Harbin Medical University, Harbin, 150001, China

    Li-yu Jiang, Xiao-lei Zhang, Ping Du & Jian-hua Zheng

Authors
  1. Li-yu Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiao-lei Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ping Du
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jian-hua Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jian-hua Zheng.

Additional information

This work was supported by a grant from the Heilongjang Province Science and Technology Commission of China (No. GB07C32304)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Jiang, Ly., Zhang, Xl., Du, P. et al. γ-Secretase inhibitor, DAPT inhibits self-renewal and stemness maintenance of ovarian cancer stem-like cells in vitro . Chin. J. Cancer Res. 23, 140 (2011). https://doi.org/10.1007/s11670-011-0140-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11670-011-0140-1

Keywords

  • Ovarian cancer stem cells (OCSCs)
  • Notch signaling pathway
  • γ-secretase inhibitor