Exogenous cripto-1 suppresses self-renewal of cancer stem cell model

Md Jahangir Alam, Ryota Takahashi, Said M. Afify, Aung Ko Ko Oo, Kazuki Kumon, Hend M. Nawara, Aprilliana Cahya Khayrani, Juan Du, Maram H. Zahra, Akimasa Seno, David S. Salomon, Masaharu Seno

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Abstract

Cripto-1 is a glycophosphatidylinositol (GPI) anchored signaling protein of epidermal growth factor (EGF)-Cripto-1-FRL1-Cryptic (CFC) family and plays a significant role in the early developmental stages and in the different types of cancer cells, epithelial to mesenchymal transition and tumor angiogenesis. Previously, we have developed cancer stem cells (miPS-LLCcm) from mouse iPSCs by culturing them in the presence of conditioned medium of Lewis Lung Carcinoma (LLC) cells for four weeks. Nodal and Cripto-1 were confirmed to be expressed in miPS-LLCcm cells by quantitative reverse transcription PCR (rt-qPCR) implying that Cr-1 was required in maintaining stemness. To investigate the biological effect of adding exogenous soluble CR-1 to the cancer stem cells, we have prepared a C-terminally truncated soluble form of recombinant human CR-1 protein (rhsfCR-1), in which the GPI anchored moiety was removed by substitution of a stop codon through site-directed mutagenesis. rhsfCR-1 effectively suppressed the proliferation and sphere forming ability of miPS-LLCcm cells in a dose-dependent manner in the range of 0 to 5 µg/mL, due to the suppression of Nodal-Cripto-1/ALK4/Smad2 signaling pathway. Frequency of sphere-forming cells was dropped from 1/40 to 1/69 by rhsfCR-1 at 1 µg/mL. Moreover, rhsfCR-1 in the range of 0 to 1 µg/mL also limited the differentiation of miPS-LLCcm cells into vascular endothelial cells probably due to the suppression of self-renewal, which should reduce the number of cells with stemness property. As demonstrated by a soluble form of exogenous Cripto-1 in this study, the efficient blockade would be an attractive way to study Cripto-1 dependent cancer stem cell properties for therapeutic application.

Original languageEnglish
Article number3345
JournalInternational journal of molecular sciences
Volume19
Issue number11
DOIs
Publication statusPublished - Nov 2018

Keywords

  • Cancer stem cells
  • Cripto-1
  • MiPS-LLCcm
  • Mouse iPS
  • Recombinant Cripto-1
  • Self-renewal

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

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  • Cite this

    Alam, M. J., Takahashi, R., Afify, S. M., Oo, A. K. K., Kumon, K., Nawara, H. M., Khayrani, A. C., Du, J., Zahra, M. H., Seno, A., Salomon, D. S., & Seno, M. (2018). Exogenous cripto-1 suppresses self-renewal of cancer stem cell model. International journal of molecular sciences, 19(11), [3345]. https://doi.org/10.3390/ijms19113345